MagPurix Extraction Kit Handbook -...

www.ZINEXTS.com Vr.5.1

MagPurix Extraction Kit Handbook

For automated purification of nucleic acids using the MagPurix 12 System

1. ZP02001 MagPurix Blood DNA Extraction Kit 200

2. ZP02002 MagPurix Blood DNA Extraction Kit 1200

3. ZP02003 MagPurix Viral Nucleic Acid Extraction Kit

4. ZP02004 MagPurix Tissue DNA Extraction Kit

5. ZP02005 MagPurix Cultured Cell DNA Extraction Kit

6. ZP02006 MagPurix Bacterial DNA Extraction Kit

7. ZP02007 MagPurix HPV DNA Extraction Kit for Swab Samples

8. ZP02008 MagPurix TB DNA Extraction Kit

9. ZP02009 MagPurix FFPE DNA Extraction Kit

10. ZP02010 MagPurix Forensic DNA Extraction Kit

11. ZP02011 MagPurix Viral/Pathogen Nucleic Acids Extraction Kit A

12. ZP02012 MagPurix Viral/Pathogen Nucleic Acids Extraction Kit B

13. ZP02013 MagPurix Viral RNA Extraction kit

14. ZP02014 MagPurix Plant DNA Extraction Kit

15. ZP02015 MagPurix Total RNA Extraction Kit

16. ZP02016 MagPurix Viral Nucleic Acid Extraction Kit 800

For in vitro diagnostic use

Version: 5.1

Rev. Date: 01 JAN, 2014

Manual No. Z3001

Welkang Limited Suite B, 29 Harley Street

LONDON, W1G 9QR,

U.K.

ZINEXTS LIFE SCIENCE CORP.

2F.-2, NO.122, Qiaohe Rd., Zhonghe

Dist., New Taipei City 235, Taiwan

(R.O.C.)

TEL: +886 2 22463579

FAX: +886 2 22438570


TABLE OF CONTENTS

Page

REAGENT KIT SELECTION GUIDE 3

INTRODUCTION 4

The Zinexts Nucleic Acid Purification Technology 4

PRODUCT INFORMATION 5

Product Use Limitation, Warranty, Guarantee and Technical Support 5 Safety Info. , Manufacturer Info 6 ZP02001 MagPurix Blood DNA Extraction Kit 200 7 ZP02002 MagPurix Blood DNA Extraction Kit 1200 15 ZP02003 MagPurix Viral Nucleic Acid Extraction Kit 20 ZP02004 MagPurix Tissue DNA Extraction Kit 26 ZP02005 MagPurix Cultured Cell DNA Extraction Kit 35 ZP02006 MagPurix Bacterial DNA Extraction Kit 40 ZP02007 MagPurix HPV DNA Extraction Kit for Swab Samples 49 ZP02008 MagPurix TB DNA Extraction Kit 54 ZP02009 MagPurix FFPE DNA Extraction Kit 60 ZP02010 MagPurix Forensic DNA Extraction Kit 66 ZP02011 MagPurix Viral/Pathogen Nucleic Acids Extraction Kit A 73 ZP02012 MagPurix Viral/Pathogen Nucleic Acids Extraction Kit B 78 ZP02013 MagPurix Viral RNA Extraction kit

ZP02014 MagPurix Plant DNA Extraction Kit

ZP02015 MagPurix Total RNA Extraction Kit

ZP02016 MagPurix Viral Nucleic Acid Extraction Kit 800

83

88

93

101

PROTOCOL OF EXTRACTION 105


Reagent Kit Selection Guide


Introduction The Zinexts Nucleic Acid Purification Technology

Introduction

Zinexts Life Science is specialized in developing advanced,

efficient and reliable technologies in nucleic acid purification,

enabling successful delivery of extraction results from varied

sample types.

The MagPurix technology is a state of the art platform that

uses magnetic bead to extract nucleic acids from samples.

The platform commits a truly walk-away automation in

nucleic acid purification from samples to results. The

purification process contains steps of lysis, binding, washing

and elution as figure below.


Product information

Intended use

MagPurix Kits is intended for the purification of nucleic acid

from biological specimen used with MagPurix 12s instrument

as IVD accessory.

The nucleic acids purified using the MagPurix 12s instrument

and reagent kits are suitable for a variety of polymerase chain

reaction (PCR) tests for human in vitro diagnostics purpose.

The MagPurix 12 instrument and reagent kits is intended for

professional use only.

Product Use

Limitations

The MagPurix instrument and MagPurix kit are not intended

for use as part of a specific in vitro diagnostic test. The user is

responsible for establishing performance characteristics

necessary for downstream diagnostic applications.

Appropriate controls must be included in any downstream

diagnostic applications using nucleic acid purified using the

MagPurix instrument and MagPurix reagent kits.

Warranty

Zinexts is committed to providing our customers with

high-quality products and services. Our goal is to ensure that

every customer is 100% satisfied with our products and our

service. If you have any questions or concerns about our

product or service, contact our Technical Support

Representatives.

Zinexts guarantees the performance of all products according

to specifications stated on our product literature. The

purchaser/user must determine the suitability of the product

for its particular use. We reserve the right to change, alter, or

modify any product to enhance its performance and design.


This warranty limits Zinexts Life Science Corporation’s liability

only to the cost of the product. No warranty is granted for

products beyond their listed expiration date. No warranty is

applicable unless all product components are stored in

accordance with instructions.

Satisfaction

Guarantee

For any product that fails to perform satisfactorily due to any

reason other than misuse, Zinexts will replace it free of

charge. Simply call your distributor to get a replacement.

Technical

Support

For technical assistance and more information, please visit

our website at www.zinexts.com or call one of the Zinexts

Technical Service Departments or local distributors.

Safety

Information

When working with chemicals or samples, always wear a

suitable lab coat, disposable gloves, and protective goggles.

For more information, please consult the appropriate material

safety data sheets (MSDSs). You can find, download, view, and

print them from our website www.zinexts.com

Manufacturer

Information

Manufacturer:

Zinexts Life Science Corp.

Address:

2F.-2, No.122, Qiaohe Rd., Zhonghe Dist.,New Taipei City 235,

Taiwan

Tel: +886 2 2246 3579

Fax: +886 2 2243 8570

Mail: [email protected]

Product of Origin: TAIWAN

http://www.zinexts.com/

mailto:[email protected]


Kit Contents ZP02001-36 ZP02001-48

Reagent Cartridge 36 pcs (6x6) 48 pcs (4x6x2)

Reaction Chamber 36 pcs (6x6) 48 pcs (4x6x2)

Tip Holder 36 pcs 48 pcs(24x2)

Filtered Tip 38 pcs 50 pcs(25x2)

Piercing Pin 38 pcs 50 pcs(25x2)

Sample Tube (2 mL) 38 pcs 50 pcs(25x2)

Elute Tube (1.5 mL) 38 pcs 50 pcs(25x2)

Barcode Paper 1 pc 1 pc

MagPurix Blood DNA Extraction Kit 200

Cat. No. ZP02001 Process Time: 50-55 minutes

Intended Use

MagPurix Blood DNA Extraction Kit is used with the

MagPurix 12 instrument for extraction of DNA from

100-400μl mammalian whole blood, suspension of

mammalian blood cells

Application

Nucleic acids extracted from MagPurix Blood DNA Extraction

kit can be used in a number of downstream application

including: PCR, qPCR, Sequencing(NGS), Microarray, RFLP,

Southern Blot Analysis

Number Of Tests 36 or 48 extractions

Kit Components


Reagent Cartridge Content

well-1 Proteinase K solution 40 μl

well-2 Lysis Buffer 2 1000 μl

well-3 Binding Buffer 1 600 μl

well-4 Magnetic Bead Solution 800 μl

well-5 Washing Buffer 1 1000 μl



well-8 Elution Buffer 1 1000 μl


well-10 Empty

Storage

MagPurix Blood DNA Extraction Kit should be

stored at room temperature (15-25°C). Do not

freeze the reagent cartridges. The Kits are stable

for 12 months under the condition

Store the purified total nucleic acid at 4 °C

(short-term) or aliquot and store at –70°C

(long-term) before perform the downstream

analysis


Starting Material

Sample type Whole blood, Buffy coat, Leukocyte concentration*

Target nucleic acid Total DNA (Genomic DNA, mitochondrial and/or viral

DNA)**

Sample volume 100-400l whole blood (WBC count less than 2x 104 cells/l) ;

100-400l leukocyte concentration (contain no more than 5x 106 cells) ;

100-400l buffy coat**

NOTE:

* For those samples which have low leukocyte count(less than 1x103

cells/l), concentrate the blood cells by centrifuge at 3000 r.p.m. for

15min under 4°C and take the leukocyte concentration for DNA

extraction is recommended

** If the WBCs no. of blood sample were more than 2 x104

cells/l,

using MagPurix blood DNA extraction kit 1200 or dilute the blood

sample with PBS is recommended (e.g. the whole blood sample from

lymphoma/myeloma patient or other granulocyte-rich blood

sample/buffy coat)

Controls/Optional

internal control#

Add controls /internal control in the extraction procedure if the

downstream analysis needed (# see Controls/internal control on page

14 )

Elute volume 50-300l

If the sample volume is less, add the appropriate

volume of PBS

MagPurix Blood DNA Extraction Kit has proven to work

for fresh or frozen blood samples collected in tubes

containing common anti-coagulants like EDTA, heparin*

and citrate (*The EDTA is recommended to use as

anticoagulation agent, while heparin have inhibit effects

on nucleic acid amplification reaction)

Using fresh whole blood sample (within 1 week) for

extraction is recommended, the total nucleic acid yield

and quality would be decreased by time

Using the concentrated buffy coat (purified and free of

blood cells), the MagPurix Culture Cell DNA extraction

kit (ZP02005) is recommended


If the whole blood sample is granulocyte-rich (white

blood cell no. more than 2 X 105 cells/l), dilute blood

sample or extract the DNA by the MagPurix blood DNA

extraction kit 1200(ZP02002) is recommended

For short-term storage (up to 10 days), store the tubes

at 2–8°C. However, for applications requiring maximum

fragment size, such as Southern blotting, storage

purified DNA at 2–8°C for up to 3 days , as low levels of

DNA degradation will occur after this time

For long-term storage, store the tubes at –70°C.

In fact, the extracted product contains total nucleic acid

(DNA and RNA), the RNA is not the major product in

this kit (about 10%) and would be degraded soon. If the

RNA-free product is needed, add some RNase to the

eluate

Result

(1) Expected Purity and Yield DNA was purified by using the MagPurix Blood DNA extraction Kit 200

and the MagPurix Purification Instrument using five different blood samples.

The DNA concentration was measured using a NanoDrop® 2000

spectrophotometer . The range of DNA yield is 2-18g ( from the sample of

WBCs counts : 2-20x103 cells/l)

Sample material Volume/amount DNA Yield Purity

Whole Blood

(WBC no. is 1.8 x 103 /ml)

100l

200l

300l

400l

1-1.2g

2-2.1g

2.8 -3.1g

4 -4.3g

OD260/OD280 >=1.7

OD260/OD230 >=1.5

Whole Blood

(WBC no. is 4 x 103 /l)

100l

200l

300l

400l

1.6 - 2.1g

3.8 - 3.9g

5.1 - 5.2g

8.5- 8.8g


Whole Blood

(WBC no. is 6.9 x 103

/l)

100l

200l

300l

400l

2.9 - 3.1g

5.8- 6.2g

8.2 -8.8g

11.9 - 12.5g

Whole Blood

(WBC no. is 10.9 x 103 /l)

100l

200l

300l

400l

4.3- 4.6g

8.7-9.1g

11.5-12.2g

16.6- 17.5g

K562 cells 6 x 105

cells

2 x 106 cells

9-9.6g

22-25g

(2) Integrity

The DNA was isolated in replicates of 12 from 200l human whole blood

taken from two different donors. Elution volume was set to 100l. Integrity

of DNA was shown by subjecting each eluate to TAE agarose gel

electrophoresis together with a suitable molecular weight marker. All

samples shown single band with molecular weight at least 20kb with no

smear

(3) Scalability A. The DNA was isolated from different whole blood samples (WBCs

count range 1.8 – 22 X 103 cells/l). The DNA yield (measured by

Nanodrop 2000 UV-Vis spectrophotometer) shows excellent scalability

in different volume (100, 200, 300 and 400l) extraction and WBCs

count of the samples


15

16

17

18

19

20

21

0 100 200 300 400 500

Sample volume (l)

Cro

ssin

g p

oin

t (C

t)

B. DNA isolated from different amounts of whole blood samples was

amplified by real-time qPCR by using theβ -globin gene specific

primers. The resulting crossing points confirm the scalability on

extraction

R² = 0.9959

R² = 0.9997

R² = 0.9981

R² = 0.9913

0

50

100

150

200

250

300

350

0 5 10 15 20 25

100ul

200ul

300ul

400ul

white blood no. ( E+3/l )

DN

A y

ield

(

l)

R² = 0.9935

0

20

40

60

80

100

120

140

0 100 200 300 400 500

Sample volume (l)

DN

A y

ield

(

g)


(4) Reproducibility

The DNA were isolated by MagPurix blood DNA extraction kit from twenty

whole blood samples. The β-globin gene was detected by real-time qPCR. This data

shows ultra high stability and reproducibility of MagPurix 12s nucleic acid

purification system.

(5) Stability The DNA were extracted from whole blood by MagPurix Blood DNA Extraction

kit 200. Real-time qPCR detection of the β-globin (A) and spiked parvovirus B-19

DNA (A, B). No significant influences on samples with different anticoagulants and

storage conditions

(6) Sensitivity Using whole blood (in EDTA collection tube) spiked with serial-diluted human

Parvo B19 Virus (in range of 25-2500000 IU/ml). 200l sample were extracted and

eluted in 100l. 10l eluate was used for real-time PCR reaction by RealStar®

Parvovirus B19 PCR kit 1.0. As little of 5 IU spiked (about 1 IU in PCR reaction)

sample can be detected, proving the excellent sensitivity and linearity of isolation

procedure


Controls/ internal control

Using appropriate controls for downstream analysis: :

Type Description Location

Positive control Using sample which positive for

target

Place in sample tube

Negative control Using sample which negative for

target or water(NTC)


Internal control (IC) Using a defined quantity control Place in sample tube or the round

well of the reaction chamber

Quality Control

In accordance with ZINEXTS’s ISO-certified Quality Management System, each lot of

MagPurix Blood DNA Extraction Kits is tested against predetermined specifications

to ensure consistent product quality

15

17

19

21

23

25

27

29

31

1 10 100 1000 10000 100000 1000000

Cro

ssin

g p

oin

t (C

t)

Human Parvo B19 virus DNA (IU/l)


MagPurix Blood DNA Extraction Kit 1200


Intended Use

MagPurix Blood DNA Extraction Kit is used with the

MagPurix 12 instrument for extraction of gDNA from

400-1000μl mammalian blood, suspension of mammalian

blood cells.

Application

Nucleic acids extracted from MagPurix Blood DNA

Extraction kit can be used in a number of downstream

application including: PCR, qPCR, Sequencing(NGS),

Microarray, RFLP, Southern Blot Analysis

Number Of Tests

36 or 48 extractions

Kit Components




















well-10 Empty


Storage

MagPurix Blood DNA Extraction Kit should be

stored at room temperature (15-25°C). Do not

freeze the reagent cartridges. The Kits are

stable for 12 months under the condition

Store the purified total nucleic acid at 4 °C

(short- term) or aliquot and store at -70°C

(long-term) before perform the downstream

analysis


Starting Material Sample type Large volume whole blood sample, Buffy coat

Target nucleic acid Total DNA (Genomic DNA, mitochondrial and viral DNA)*

Sample volume 400-1000l whole blood (WBC count less than 6x 104cells/l) ;

400-1000l buffy coat*

NOTE:

*If the WBC no. of whole blood sample were more than 6 x104

cells/l,

dilute the sample with PBS or saline is recommended

Controls/Optional

internal control#


downstream analysis needed (# see Controls/internal control on page 18 )

Elute volume 100-400l*

*For those WBCs no. > 30000/l samples, set elute volume more than

200l is recommended

MagPurix Blood DNA Extraction Kit has proven to work

for fresh or frozen blood samples collected in tubes

containing common anti-coagulants like EDTA, heparin*

and citrate(*The EDTA is recommended to use as

anticoagulation agent, while heparin have inhibit effects

on nucleic acid amplification reaction)

If the sample volume is less than 400l or the WBCs no.

is high (more than 60000 cells/l ), add the appropriate

volume of PBS to adjust

Using fresh whole blood sample (within 1 week) for

extraction is recommended, the total nucleic acid yield

would be decreased by time

For short-term storage (up to 10 days), store the tubes

at 2–8°C. However, for applications requiring maximum

fragment size, such as Southern blotting, storage

purified DNA at 2–8°C for up to 3 days , as low levels of

DNA degradation will occur after this time

For long-term storage, store the purified DNA at –70°C.

In fact, the eluate contains total nucleic acid (DNA and

RNA), the RNA is not the major product in this kit (about

10%) and would be degraded soon. If the RNA-free

product is needed, add some RNase to the eluate


Expected Purity and Yield

DNA was purified by using the MagPurix Blood DNA extraction Kit 1200

and the MagPurix Purification Instrument using four different blood samples.

The DNA concentration was measured using a NanoDrop® 2000

spectrophotometer. The range of DNA yield is 2-60g (in the WBCs range 2-60

x 103cells/l)

Sample material Volume/

amount

DNA Yield Purity

Whole Blood

(WBC no. is 6 x 103 /ml)

400l

1000l

9-9.5g

25-30g

OD260/OD280 >=1.7

OD260/OD230 >=1.5

Whole Blood

(WBC no. is 11 x 103 /l)

400l

1000l

18-19g

45-50g

Whole Blood

(WBC no. is 25x 103 /l)

400l

1000l

30-36g

70-75g

Whole Blood

(WBC no. is 40 x 103 /l)

1000l 100g*

Buffy coat 100l

200l

10-30g

20-60g

* For those WBCs count > 30000/l samples, set elute volume more than 200l is

recommended





target








Quality Control

In accordance with ZINEXTS’s ISO-certified Quality Management System, each

lot of MagPurix Blood DNA Extraction Kits is tested against predetermined

specifications to ensure consistent product quality


MagPurix Viral Nucleic Acid Extraction Kit


Intended Use

MagPurix Viral Nucleic Acid Extraction Kit is used with the

MagPurix 12 instrument for extraction of Viral DNA or RNA

from human biological specimens such as serum, plasma,

and other cell-free fluids

Application

Nucleic acids extracted from MagPurix Viral Nucleic Acid




Number Of Tests


Kit Components




Tip Holder 36 pcs 48 pcs (24x2)

Filtered Tip 38 pcs 50 pcs (25x2)

Piercing Pin 38 pcs 50 pcs (25x2)

Sample Tube (2 mL) 38 pcs 50 pcs (25x2)

Elute Tube (1.5 mL) 38 pcs 50 pcs (25x2)

RNA Carrier (1mg) 1 pc 1 pc











well-8 RNase-free water 1000 μl

well-9 Rnase-free water 1000 μl

well-10 Empty

Storage

MagPurix Viral Nucleic Acid Extraction Kit should be

stored at room temperature (15-25°C). Do not freeze

the reagent cartridges. The Kits are stable for 12

months under the condition

After dissolve the carrier RNA, store it at 4°C

(short-term, up to 1 month) or -20°C (long-term). Do

not freeze–thaw the Frozen carrier RNA more than 3

times


Starting Material

Sample Type Target Nucleic Acid

Sample Volume (Amount of starting material)

Elution Volume

Serum Total Viral

Nucleic Acids

(DNA + RNA)

100-400μl 50-300μl Plasma

CSF

Pretreated Urine

Cell-free body

fluids

Controls/internal

control#

Add controls /internal control in the extraction procedure if

the downstream analysis needed (# see control/internal

control on page 24 )

The kit is designed for extraction of viral nucleic

acids (e.g., those of HIV, HCV, HBV, CMV and

EBV) from plasma or serum, or from a pool of

such cell-free body fluids

After extraction, store the nucleic acid at 4°C

(up to 24hours) or -20°C for longer storage.

Repeated freeze–thawing is not allowed

Sample preparation

The purification procedure is optimized for use

with 100- 400 μl serum, plasma, CSF, or

pretreated urine samples. ( Blood samples

treated with EDTA or citrate as an anticoagulant

can be used for plasma preparation)

Samples can be either fresh or frozen, provided

that they have not been refrozen after thawing

After collection and centrifugation, plasma,

serum, or CSF can be stored at 2–8°C for up to 6

hours. For longer storage, we recommend

freezing aliquots at –20°C or –80°C. Thaw

samples at room temperature (15–25°C), and


process the samples immediately when they

have equilibrated to room temperature. Do not

refreeze the aliquots after thawing. Repeated

freeze–thawing leads to denaturation and

precipitation of proteins, resulting in reduced

viral titers and therefore reduced yields of viral

nucleic acids. If cryoprecipitates are visible in

the samples, centrifuge at 6800 x g for 3

minutes, transfer the supernatants to fresh

tubes without disturbing the pellets, and start

the purification procedure immediately

Carrier RNA For RNA virus, adding carrier RNA to the

sample before extraction is recommended!!!

Add 1.0 ml RNase - free water to the carrier

RNA tube (provided with the kit) and mix by

vortexing. Store it at 4°C (short-term, up to 1

month) or -20°C (long-term). Do not

freeze–thaw the Frozen carrier RNA more than

three times

Add 5 μl Carrier RNA (for 100 μl sample), 10 μl

(for 200 μl sample) or 20 μl (for 400 μl sample)

into the Sample Tube


Result

(a) DNA virus (HBV)

Using serum spiked with serial-diluted Hepatitis B Virus (in range of

300000-30 IU/ml). 200l sample were extracted and eluted in 100l. 30l

eluate was used for real-time PCR reaction by AmpliSens®

HCV/HBV/HIV-FRT PCR kit. As little of less than 6 IU spiked (about 1 IU in

PCR reaction) sample can be detected, proving the excellent sensitivity

and linearity of isolation procedure

(b) RNA virus (HCV)

Using serum spiked with serial-diluted Hepatitis C Virus (in range of

500000-50 IU/ml). 200l sample were extracted and eluted in 100l. 30l

eluate was used for real-time PCR reaction by AmpliSens®

HCV/HBV/HIV-FRT PCR kit. As little of less than 10 IU spiked (about 3 IU in

PCR reaction) sample can be detected, proving the excellent sensitivity and

linearity of isolation procedure



Using appropriate controls for downstream analysis:



target





Internal control (IC) Using a defined quantity control Place in sample tube or the round well of

the reaction chamber

Quality Control


MagPurix Viral Nucleic Acid Extraction Kits is tested against predetermined specifications

to ensure consistent product quality


MagPurix Tissue DNA Extraction Kit


Intended Use

MagPurix Tissue DNA Extraction Kit is used with the MagPurix 12 instrument for extraction of genomic DNA from a variety of animal tissues, swab samples and blood stain

For extraction from FFPE samples, using “ZP02009 MagPurix

FFPE DNA extraction kit” is recommended

Application

Nucleic acids extracted from MagPurix Tissue DNA Extraction kit

can be used in a number of downstream application including:

PCR, qPCR, Sequencing(NGS), Microarray, RFLP, Southern Blot

Analysis

Number Of

Tests


Kit Components









Proteinase K (10mg/mL)

1 pc (1mL) 1 pc (1mL)

BL2 Buffer 1 pc (25mL) 1 pc (25mL)




well-1 Empty









well-10 Empty

Storage

MagPurix Tissue DNA Extraction Kit should be stored at room

temperature (15-25°C). Do not freeze the reagent cartridges.

The Kits are stable for 12 months under the condition

Store the purified DNA at 4 °C (short- term) or aliquot and

store at -70°C (long-term) before perform the downstream

analysis


Starting

Material

The types and amounts of starting material for use in MagPurix

Tissue DNA purification procedures are shown in Table listed

below,

Sample Type Target

Nucleic Acid

Sample Volume

(Amount of starting material)

Elution

Volume

Tissue DNA 100-400 μl/10-40 mg 50-300μl

Dried swab samples (e.g. Buccal cells)

100-400 μl/1 swab or brush

(add BL2 and proteinase K to

100-400l for extraction)

Dried blood 100-400 μl/4 discs*

Control/Optional

internal control#


downstream analysis needed (# see control/internal control on page

33 )

*A 3mm diameter disc punched out from filter paper stained with dried

blood contains white blood cells from approximately 5 μl whole blood; we

recommend using 4 punched-out discs as starting material

Yield of

purified DNA

DNA yields depend on the sample type, number of nucleated

cells in the sample, and the protocol used for purification of

DNA

Table listed below shows DNA yields obtained from different

sample types using MagPurix extraction procedures


Table: The DNA yield of different sample types

Sample Type Sample Amount Typical DNA Yield

Skeletal muscle 200 μl

(40 mg tissue digested)

Up to 9μg

Heart 200 μl


Up to 12μg

Spleen 200 μl


Up to 27μg

Lung 200 μl


Up to 17μg

Kidney 200 μl


Up to 18μg

Liver 200 μl


Up to 40μg

Buccal cells 1 Swab 1-5 μg

Duried blood 4 x 3 mm diameter discs 0.2-0.5 μg

Sample preparation requirements are highly dependent upon the type of

starting material. Due to variations in consistency and viscosity, even similar

sample types may require distinct handling. The steps below describe some

recommendations for processing primary samples


Sample preparation

For solid animal tissue

��1. �Transfer tissue

No. Sample

Type

Recommended

Sample Amount

1 Heart 20 mg

2 Muscle 40 mg

3 Other

Tissues

10 mg

2. �Add BL2 Buffer

�3. Add Proteinase K

4. �Incubate

5. �Spin down and

Transfer

Spin down and transfer 100-400μl the

supernatant to Sample Tube. Use BL2 buffer to

adjust the sample volume.

OPTIONAL: Use the filter column

(ZA030118, not supplied in kit) to remove

the residual debris and mucous material

before DNA extraction will increase the

DNA yield (20-100%)

Incubate at 55°C in a shaking water bath or

thermomixer (mix set at 1000r.p.m.)until the

tissue is completely lysed.

Note: 1. Lysis time varies depending on the

type of tissue processed. Lysis is usually

completed in 1-2 hrs. However, lysis overnight is

possible and does not influence the preparation.

2. Use the heat block for incubation, vortexing-

mix several times during incubation is

recommend

Transfer tissue into a 1.5ml microcentrifuge

tube:

Add 100-400 μl provided Buffer BL2. Ensure

tissue pieces are fully submerged in Buffer BL2

Add 20 μl proteinase K solution and mix by

vortexing


For Swap Tissue

�1. Cut�� Carefully cut or break off the end part of

the swab or brush into a 2 ml screw-

capped tube (not supplied) using an

appropriate tool (e.g., scissors).

��2. Add BL2 Buffer Add 100-400 μl of Buffer BL2 to the

sample.Ensure sample is fully submerged

in Buffer BL2

�3. Add proteinase K Add 20 μl proteinase K, and mix

thoroughly by vortexing for 10 s.

If processing buccal cell brush samples,

centrifuge the tube briefly (at 10,000 x g

for 30s) to force the brush to the bottom

of the tube.

4. ��Incubate Incubate at 55°C for 15 min (place in a

thermomixer, mix set at 1000 r.p.m. or

votex mix several times during incubation

in heat block)�

�5. Centrifuge Centrifuge the tube briefly to remove

drops from inside the lid.

�6. Remove Remove the swab or brush from the

tube.Using forceps, press the swab or

brush against the inside of the tube to

obtain maximum sample volume. The

sample volume should be approximately

as the setting volume(100-400l). *Use

BL2 buffer to adjust the volume


For FFPE Tissue

To extract DNA from FFPE samples, please select and refer to

MagPurix FFPE DNA Extraction Kit (ZP02009)


Dried Blood





target







1.� Collect Collect 70 μl of each blood sample

onto a ring marked on filter paper.

Allow the blood to air dry.

Either untreated blood or blood

containing anticoagulant (EDTA, ACD,

or heparin) can be used.

2.� Cut For each dried blood sample, use the

manual paper punch to cut out four 3

mm diameter discs.

3.� Add BL2 Transfer each set of 4 discs to a 1.5 ml

microcentrifuge tube. Add 220-440l

BL2 to sample

4.� Add

proteinase K

Add 20l proteinase K and mix by

vortexing

5.� Incubation incubate at 55℃, 15min in a

thermomixer (set mix at 1000r.p.m.)

or vortex mix several time during

incubation in heat block

6.� Centrifuge Centrifuge the tube brifly to remove

the drop inside the lid

7.� Transfer Transfer the 100-400l supernatant to

the sample tube (provided in the kit),

proceeding DNA extraction


Quality Control


MagPurix Tissue DNA Extraction Kit is tested against predetermined specifications to

ensure consistent product quality.


MagPurix Cultured Cell DNA Extraction Kit

Cat. No. ZP02005 Process Time: 40- 45 minutes

Intended Use

MagPurix Cultured Cell DNA Extraction Kit is used with the

MagPurix 12 instrument for extraction of genomic DNA

from culture cells and buffy coat

Application

Nucleic acids extracted from Cultured Cell DNA Extraction




Number Of

Tests


Kit

Components





















well-10 Empty

Storage

MagPurix Cultured Cell DNA Extraction Kit should be

stored at room temperature (15-25 °C). Do not freeze



Store the purified DNA at 4 °C (short- term) or aliquot

and store at -70°C (long-term) before perform the

downstream analysis


Starting

Material

1. Culture cells in suspension and monolayer

2. Cells from Buffy coat (without red blood cells)

If the buffy coat is remove directly from the whold

blood, for completely remove and lyse red blood cell,

the MagPurix Blood DNA Extraction kit (ZP02001;

ZP02002) is recommended

Do not use more than 5 x 106 cells with a normal set

of chromosomes

The cell number can be determined by using

Hemocytometer1,2 (Petroff-Hauser Chamber )and

automated cell counter( e.g. TC10™, Countess®,

Cellometer®, and Scepter™ automated cell

counters )

The types and amounts of starting material for use in

MagPurix Cultured Cell DNA purification procedures

are shown in Table listed below

Reference

1. http://www.smccd.edu/accounts/case/biol230/algae/hemo

cytometer1.pdf

2. http://web.mnstate.edu/provost/CountingCellsHemocytom

eter.pdf

Sample preparation

Cells grown in suspension

Suspension culture:

1. Determine the number of cells. ( Do not use

more than 5 x 106 cells with a normal set of

chromosomes)

2. Centrifuge the appropriate number of cells for 5

min at 1000 x g in a 1.5 ml microcentrifuge tube

3. Remove the supernatant completely and discard,

taking care not to disturb the cell pellet

4. Resuspend cell pellet in PBS to a final volume of

200 μl

http://www.smccd.edu/accounts/case/biol230/algae/hemocytometer1.pdf

http://www.smccd.edu/accounts/case/biol230/algae/hemocytometer1.pdf

http://web.mnstate.edu/provost/CountingCellsHemocytometer.pdf

http://web.mnstate.edu/provost/CountingCellsHemocytometer.pdf


Cells grown in a monolayer

Cells grown in a monolayer can be detached from

the culture flask by either trypsinization or using a

cell scraper

To trypsinize cells:

1. Determine the number of cells* (do not use more

than 5 x 106 cells with a normal set of

chromosomes)

2. Aspirate the medium and wash cells with PBS

3. Aspirate the PBS, and add 0.10–0.25% trypsin,

incubate at 37°C

4. After cells have detached from the dish or flask,

collect them in medium, and transfer the

appropriate number of cells to a 1.5 ml

microcentrifuge tube.

5. Centrifuge for 5 min at 1000 x g.


taking care not to disturb the cell pellet.


200 μl.

Using a cell scraper:

1. Determine the number of cells. (do not use more

than 5 x 106 cells with a normal set of

chromosomes)

2. Detach cells from the dish or flask by scraping

3. Harvest and transfer cells to a 1.5 ml

microcentrifuge tube and centrifuge for 5 min at

1000 x g.


taking care not to disturb the cell pellet.


200 μl


Yield Of

Purified DNA

DNA yields depend on the sample type, number of

nucleated cells in the sample, and the protocol used for

purification of DNA.

For example, the average DNA yield form the HT29 colon

adenocarcinoma cell line at the different concentrations

( in the range from 1 x 105 to 106 cells) is about 22g/106

cells as below

Quality Control

In accordance with ZINEXTS’s ISO-certified Quality Management System,

each lot of MagPurix Cultured Cell DNA Extraction Kits is tested against

predetermined specifications to ensure consistent product quality


MagPurix Bacterial DNA Extraction Kit


Intended Use

MagPurix Bacterial DNA Extraction Kit is used with the MagPurix 12 instrument for extraction of genomic DNA from both Gram-positive and Gram-negative bacteria

Application

Nucleic acids extracted from Bacterial DNA Extraction kit can be used in a number of downstream application including: PCR, qPCR, Sequencing(NGS), Microarray, RFLP, Southern Blot Analysis

Number Of

Tests


Kit

Components









BL2 Buffer 1 pc (25mL) 1 pcs (25mL)













well-10 Empty

Storage

MagPurix Bacterial DNA Extraction Kit should be stored at

room temperature (15-25°C). Do not freeze the reagent

cartridges.The Kits are stable for 12 months under the

condition


store at -70°C (long-term) before perform the downstream

analysis


Starting

Material

Bacterial pellet/colony from culture, cell-free body fluids,

liquid transport media, urine, environment material (water, soil, etc.)

Use the paraffin- embedded tissue sections as samples, we recommend to extract DNA by MagPurix FFPE DNA Extraction kit (ZP02009)

Use tissue as samples, we recommended use the MagPurix Tissue DNA Extraction kit


MagPurix Bacterial DNA purification procedures are shown in Table listed below,


Sample volume (Amount of starting material)

Elution Volume

Bacteria Pellet Genomic DNA 200-400 μl /Up to 109

bacteria (about OD600 = 3)

50-300 μl

Bacterial colony 200-400 μl /1-3 colony

Tissue 200-400 μl /1-30 mg

Urine 200-400 μl /5-50 mL urine

Cell-free body fluids

200-400 μl cell-free body fluids

Liquid transport media

200-400 μl liquid transport media

NOTE: Before extraction, adjust sample volume with BL2 buffer

Sample

Preparation

Sample preparation requirements are highly dependent

upon the type of starting material. Due to variations in consistency and viscosity, even similar sample types may require distinct handling

The buffer BL2 is specialized for bacterial cell wall lyse* (Supplied in the kit), use it to resuspend the bacterial pellet before process extraction

* For mycobacterium spp.(e.g. MTB), use buffer BL3 for


Table : Preparation of sample material for bacterial nucleic acid extraction

Sample type Procedure

For viscous samples e.g. BAL, sputum or other mucous specimen

Recommended pretreatment : Liquefaction 1. Prepare a fresh DTT stock solution for

liquefaction * (e.g., 5× conc. DTT stock is about 0.75%)

2. Adjust the final DTT concentration in the sample

to 0.15% by adding DTT stock solution.

3. Incubate the sample (e.g., with shaking at 850r.p.m. for 30 min at 37°C) until it can be pipette easily.

4. Pellet bacteria by centrifugation at 14000 x g for 10 min

5. Discard supernatant, resuspend the pellet in 220 μL Buffer BL2

6. Transfer 200 μL suspension to sample tube (Supplied in the kit) * The liquefaction could be done by using other solutions, such as NALC（N-Acetyl-L-Cysteine）

-NaOH or other agents which could digest mucous material

bacterial cell wall lysis (BL3 buffer is supplied in the ZP02008

MagPurix TB DNA Extraction kit )

The table below describes the recommendations in processing the primary samples before nucleic acid extraction:


For large volume liquid samples that have low or unknown bacterial loads e.g. urine, water collected from pool/river stream/tower

Recommended pretreatment : Centrifugation 1. Centrifuge the sample for up to 10 min at

20,000 × g to concentrate the bacterial cells in pellet

2. Discard supernatant, resuspend the pellet in 220 μL Buffer BL2*

3. Take 200 μL suspension to sample tube (Supplied in the kit)

* If there were sand or other visible particle in the pellet, centrifuge again after BL2 buffer treatment or filter out the dust is recommended

For cell-free body fluids (e.g. CSF, BAL, aspirates)

Recommended pretreatment : Centrifugation Method 1 1. Pellet bacteria by centrifugation at 14000 x g

for 10 min 2. Resuspend bacterial pellet in 220 μL Buffer BL2 3. Take 200 μL suspension to sample tube

(Supplied in the kit) Method 2-Centrifugation free

1. Take 200 l sample in a 1.5 ml centrifuge tube

2. Add 200l buffer BL2 to sample (1:1) 3. Vortex-mixing for 5-10sec

4. Transfer 400l sample to sample tube (Supplied in the kit)

For swab samples e.g. eye, nasal, pharyngeal, or other swabs

Method 1 1. Collect samples and place in 2 ml PBS

containing a common fungicide. Incubate for 30min at room temperature

2. Pellet bacteria by centrifugation at 14000 x g for 10 min

3. Resuspend bacterial pellet in 220 μL Buffer BL2

(Supplied in the kit)



Method 2- centrifuge free

1. Place the sample swab in 440l buffer BL2, incubate for 30min at room temperature

2. Transfer 400l to sample tube

For some gram-positive bacteria species. Especially for samples that contain particles e.g. stool

Recommended pretreatment : Mechanical homogenization

Follow the regular homogenization procedures

in the laboratory.

For some sample types, DNA yield can be

improved by performing this homogenization

step prior to add buffer BL2 and proteinase K

Isolation of genomic DNA from bacterial suspension cultures

1. Pipet 1 ml of bacterial culture into a 1.5 ml

microcentrifuge tube and centrifuge at 5000xg for 5 min

2. Discard supernatant

3. Add 220l Buffer BL2 to pellet and mix by vortexing for 5-10 sec

4. Take 200 μL suspension to sample tube

(Supplied in the kit)

Isolation of genomic DNA

from bacterial plate

culture

1. Take 1-3 bacterial colony from culture plate

with an inoculation loop and suspend in 220 μL of buffer BL2 by vigorous stirring



To inactive pathogenic

organisms in the sample

Recommended pretreatment : Boiling

1. Incubate samples at 95°C for 10 min

2. Centrifuge briefly to collect the complete

sample volume at the bottom of the tube.

3. Allow samples to cool down or chill on ice,

then transfer 100-400l cooled sample to the

sample tube

Yield of purified DNA

DNA yields depend on the sample type, number of bacteria in the sample, and the protocol used for purification of DNA

Result

(1) Scalability Using MagPurix Bacterial DNA Extraction kit to isolate the DNA from

cultured Escherichia. coli (ATCC25922) and Staphylococcus aureus

(ATCC27154) in LB broth at different bacterial density (measure the Optical

Density at 600nm; OD600). Take 200l bacterial culture for extraction and

collect the eluate in 100l. The total nucleic acid yield of different bacterial

density was measured by Nanodrop 2000 UV-Vis spectrophotometer (fig.1a

and 2a) and analyzed by 1% TAE agarose gel electrophoresis (fig.1b and 2b).

The result shown the nucleic acid extraction in both Gram-negative (E.coli)

and Gram-positive (S. aureus) were have excellent scalability.

0

5

10

15

20

0 0.5 1 1.5 2 2.5

tota

l nu

cle

ic a

cid

yie

ld (

g)

Turbidity (O.D.600)

Fig.1a E. coli


Fig.1b

Fig.2b

(2) Sensitivity Performing serial-dilution on Staphylococcus aureus (ATCC27154) in range

of 109-101copy/ml). 200l sample were extracted and eluted in 100l. 25l

eluate was used for SYBR Green real-time PCR reaction which detect

Staphylococcus aureus specific gene. As little of 20 copies (about 102

copy/ml bacteria in the sample) spiked-in (about 5 copy in PCR reaction)

bacteria can be detected, proving the excellent sensitivity and linearity of

isolation procedure (fig.3a and 3b)

0

5

10

15

0 0.5 1 1.5 2 2.5 3

tota

l nu

cle

ic a

cid

yie

ld (

g)

Turbidity (O.D.600)

Fig.2a Staphylococcus aureus


Fig. 3a

Fig.3b





target







Quality Control In accordance with ZINEXTS’s ISO-certified Quality Management System,

each lot of MagPurix Bacterial DNA Extraction Kits is tested against

predetermined specifications to ensure consistent product quality


MagPurix HPV DNA Extraction Kit for

Swab samples

Cat. No. ZP02007 Process Time: 40-50minutes

Intended Use

MagPurix HPV DNA Extraction Kit is used with the MagPurix 12

instrument for DNA extraction of the Human Papillomavirus

(HPV) from cervical cell samples which collected by cervical

brush or genital swab in liquid-based Medium* (e.g. Hologic

Thinprep PreservCyt®, BD SurepathTM, etc.) or other STM

(sample transport media) preservation solutions(e.g. QIAGEN

DNA PAP Cervical sampler, Roche Cobas® PCR Cell Collection

Media, Hybribio cell preservation solution, etc.).

*The liquid-base medium are formulated for cellular

preservation and used in liquid-based cytological systems(LBC)

for cytological and molecular diagnosis

Application

Nucleic acids extracted from HPV DNA Extraction kit from swab sample can be used in a number of downstream application including: PCR, qPCR, Sequencing (NGS), Microarray, RFLP, Southern Blot Analysis, etc.

Number Of

Tests













well-10 Empty

Kit

Components









BL4 Buffer 1 pc (25mL) 1 pcs (25mL)



Storage

MagPurix HPV DNA Extraction Kit for swab

samples should be stored at room temperature

(15-25°C). Do not freeze the reagent cartridges.

The Kits are stable for 12 months under the

condition

Store the purified DNA at 4 °C (short- term) or

aliquot and store at -70°C (long-term) before

perform the downstream analysis

Starting Material

Cervical cells collected by cervical brush or

genital swab

For those non-liquid based medium, adding

BL4 Buffer to the reservation is

recommended

The specimen should be sent at 4-30°C for

examination on immediately after collection.

The storage condition is depended on the

preservation solution

Add controls /internal control in the

extraction procedure if the downstream

analysis needed (see control/internal control

on page 52 )


Sample Preparation


In liquid-based


(eg. Hologic Thinprep

PreservCyt®, BD

SurepathTM)

1. Take the sample amount as the following assay

recommended*

2. Centrifuge at 1000 x g. for 5min

3. Discard supernatant

4. Resuspend pellet in 220l BL4

5. Incubate at RT, 5 min

6. Vortex for 5 sec

7. Take 200 l suspension for extraction

In other STM


(QIAGEN DNA PAP, Hybribio

cell preservation solution)

1. Add equal volume of BL4 directly to the reservation

solution (BL4: reservation = 1:1) **

2. Incubate at RT, 5-10 min

3. Vortex for 5 sec

4. Take 100-400 l sample for extraction

* Especially for those assay of” signal amplification”, in that the target DNA won’t be

amplified. Take adequate amount of sample is necessary

** The sticky mucus is common in cervical specimen, adding BL4 before processing

will help sample liquefying and nucleic acid extraction





target





Internal control (IC) Using a defined quantity control Place in sample tube or

the round well of the

reaction chamber


Quality Control


MagPurix HPV DNA Extraction Kits for swab samples is tested against predetermined



MagPurix TB DNA Extraction Kit


Intended use

MagPurix TB DNA Extraction Kit is used with the MagPurix 12

instrument for extraction of genomic DNA of Mycobaceteria

spp. (e.g. Mycobacterium tuberculosis) from different specimen

Application

Nucleic acids extracted from TB DNA Extraction kit can be used

in a number of downstream application including: PCR, qPCR,

Sequencing(NGS), Microarray, RFLP, Southern Blot Analysis

Number Of

Tests


Kit

Components









BL3 Buffer 1 pc (25mL) 1 pc (25mL)













well-10 Buffer N1 400 μl

Storage

MagPurix TB DNA Extraction Kit should be stored

at room temperature (15-25 °C). Do not freeze






Starting Material

1. Clinical specimen: Sputum, BAL, Pas, blood,

cell-free body fluids, urine and other respiratory

specimens

2. Bacterial culture in the solid and liquid media For the MTB is highly infectious agent, prepare

sample in the BSC(Biosafety cabinet)


Sample preparation

Specimen type

Procedure

Sputum/BAL or other

Respiratory Specimen

Method 1

1. Liquify the sample*

2. Pellet bacteria by centrifugation at 12500 x g

for 15 min

3. Discard supernatant, resuspend the pellet in

200 μL Buffer BL3, votex mixing about 5 sec

4. Take 200 l sample to sample tube for

extraction

Method 2 - Centrifuge free

1. Liquify the sample*

2. Transfer 200l sample to sample tube

3. Add 200l buffer BL3 to sample (1:1)

1.

* The liquefaction could be done by using

liquefying agents, such as NALC（N-Acetyl-L-

Cysteine）- NaOH and 0.75% DTT (5x stock)

which could digest mucous material

Viscous body fluid e.g. Pas

See the procedure of “Sputum/BAL or other

Respiratory Specimen”

Cell-free body fluid

e.g. CSF, urine


for 15 min

2. Discard supernatant, resuspend bacterial

pellet in 200 μL Buffer BL3,Votex-mixing

about 5 sec

3. Take 200 μL sample to sample tube for

extraction

Liquefied,

decontaminated sample

See the procedure of “cell-free body fluid”


Blood or

Blood-contaminated

sample

1.

1. Add cold sterilized water to sample to the

ratio of water/blood about 3:1

2. Inverted mix several times

3. Incubate at 4℃, at least 10 min

4. Centrifuge at 14000 x g for 15 min

5. Remove supernatant, add 200 μL buffer BL3,

votex mixing about 5-10sec


extraction

Colony from solid

culture

1. Pick 1-3 colony, mix with 200 μL Buffer BL3,

votex mixing about 5-10 sec


extraction

Liquid culture Method 1

1. Take 1mL culture(＞McFarland 0.5), transfer

to 1.5mL microcentrifuge tube


for 5 min

3. Discard supernatant, resuspend bacterial

pellet in 200 μL Buffer BL3,Votex mixing

about 5-10 sec


extraction

Method 2- centrifuge free

1. Add Buffer BL3 to liquid culture(1:1)

2. Votex mixing about 5-10 sec

3. Take sample mixture to sample tube for

extraction






target







Result

Using MagPurix TB DNA Extraction kit to isolate DNA from clinical specimens

(sputum, CSF and pas). 100ml sample used for extraction and collect 100ml eluate.

Analysis was performed by real-time qPCR with Taqman probe /primers (IS6110).

Even in the cell-free body fluid (CSF) and blood contaminated sample(Pas), the TB

DNA can be detected after extraction, proving the excellent sensitivity of isolation

procedure

Expected Purity and Yield

DNA yields depend on the sample type, number of bacteria in the sample, and the

protocol used for purification of DNA


Quality Control

In accordance with ZINEXTS’s ISO-certified Quality Management System,

each lot of

MagPurix TB DNA Extraction Kits is tested against predetermined



MagPurix FFPE DNA Extraction Kit


Intended

Use

MagPurix FFPE DNA Extraction Kit is used with the MagPurix 12 instrument for extraction of genomic DNA from FFPE (Formalin-Fixed, Paraffin-Embedded) tissue samples. Providing good quality, high integrity DNA for Molecular diagnosis and research works

Application

Nucleic acids extracted from FFPE DNA Extraction kit can be used in a

number of downstream application including: PCR, qPCR,


Number Of

Tests


Kit

Components









Proteinase K (10mg/mL) 1 pc (1mL) 2 pcs (0.6mL,1x2)

BL2 Buffer 1pc (25 mL) 1 pcs (25mL)

Filter Column 38 pcs 50 pcs (25x2)

Collection Tube 38 pcs 50 pcs (25x2)




well-1 Empty









well-10 Empty

Storage

MagPurix FFPE DNA Extraction Kit should be stored at room

temperature (15-25 °C). Do not freeze the reagent cartridges. The Kits

are stable for 12 months under the condition

Store the purified DNA at 4 °C (short- term) or aliquot and store at

-70°C (long-term) before perform the downstream analysis


Starting

Material

FFPE (formalin fixed Paraffin Embedded) tissue samples: one to five

10m-thick section(s)

Sample Type Target

Nucleic Acid

Sample Volume

(Amount of starting material)

Elution

Volume

FFPE (formalin fixed Paraffin Embedded) tissue samples

DNA 100-400μl/ One to eight 10 μm-

thick sections (after proteinase

K digestion)*

50-300l

Needle biopsy 100-400μl/ three to ten

biopsies

*Note: The size of tissue area should be more than 1 x 1 cm2, if the area is smaller, use

more than one section for extraction.

Yield of

purified

DNA

DNA yields depend on the sample type, number of nucleated cells in

the sample, and the thickness of the section

Sample preparation requirements are highly dependent upon the

type of starting material. Due to variations in consistency and

viscosity, even similar sample types may require distinct handling.

The steps below describe some recommendations for processing

primary samples

The DNA of FFPE tissue sample is often fragmented which cause

problems in molecular assay. Keep the integrity of DNA is most

important in whole procedure.

Because of the DNA fragments often cause high OD260 and ratio of

260/280. The integrity of FFPE DNA can’t be determined only by

UV-VIS spectrophotometer analysis. The best method for the

integrity check is performing a PCR reaction of house-keeping genes

with different length products


Sample preparation

1. Transfer sections Transfer sections of paraffin-embedded tissue into a 1.5ml

microcentrifuge tube

*for most tissue types, slice of one to five 10mm-thick sections

is recommened; however, up to ten very small sections such as

needle biopsies may be used

2. Deparaffin

(Optional)

Treat FFPE samples with xylene or other deparaffin agents

*This step is optional, skip this step is still work for the

extraction

3. Add BL2 Buffer Add 400l BL2, ensure that the samples are fully submerged in

BL2 buffer

4. Add proteinase K Add 20ml Proteinase K to sample mixture, vortex mixing for

5-10 sec. short spin for remove the drop from the lid

5. Incubation Incubate sample at 55℃ for 2 hours, with vigorous mixing in a

shaking water bath or thermomixer

*The first 2 hour is the critical step for proteinase lysis.

Incubating sample with shaker is necessary (or vortex mixing

every 30 minutes) for completely digestion. Increase the

incubation time is not much helpful for lysis, because the

proteinase activity drops after 2 hours. Only when the tissue

sample is thick, add more proteinase K and incubate longer is

recommended

6. Centrifuge Centrifuge the tube briefly to remove drops from inside the lid

7. Homogenize Homogenize the sample by pipetting up and down several

times. Transfer the sample to the filter column sitting in a

collection tube (supplied in the kit). Centrifuge at 6000xg, 1min

*Pieces of insoluble material, which could clog tips, make

interferences of extraction efficiency. Remove them by filter

8. Transfer Transfer 400l sample to sample tube


Result

Integrity

Take eleven Myeloma FFPE section samples (10m thick, about 2 x 2 cm2) for extraction.

Two sections were extracted by QIAGEN FFPE Tissue kit; the others were extracted by

MagPurix FFPE DNA Extraction kit. Using DNA ladder PCR (a multiplex primer PCR of

different length products of house-keeping gene) and capillary electrophoresis to analyze

the integrity of the extracts of PCR products(fig.1)

Fig. 1 The results of capillary electrophoresis shown a quirt good integrity of the extract

Fig.1a QH and QA were the extracts from QIAGEN FFPE tissue kit (elute in water and ATE)

and extracts from MagPurix FFPE DNA were labeled as no. 1 to 9

Scalability

Using Myeloma FFPE section samples (10m thick, about 2 x 2 cm2) for extraction.

Measuring the eluate by UV-VIS Nanodrop2000 spectrophotometer

No. of section Con. (ng/l) Spectrum

1/4 section 1.5

1/2 section 3.8

1 section 8.3

2 section 15.8

3 section 24.7






target







Quality Control In accordance with ZINEXTS’s ISO-certified Quality Management System, each lot of

MagPurix FFPE DNA Extraction Kits is tested against predetermined specifications to

ensure consistent product quality


MagPurix Forensic DNA Extraction Kit


Intended

Use

MagPurix Forensic DNA extraction kit is extract and isolate

genomic DNA from forensic samples

Application

The extracted DNA is compatible for use in quantitation using

the Quantifiler® Human, Quantifiler® Y Human Male, Quantifiler®

Duo DNA Quantification Kits and Investigator® Quantiplex kit, and

for use in STR amplification using the AmpFlSTR® PCR

Amplification kits

Number Of

Tests


Kit

Components









Proteinase K (10mg/mL)

1 pc (1mL) 2 pcs (0.6mL,1x2)

Buffer BL2 1pc (25 mL) 1 pc (25mL)

Filter Column 38 pcs 50 pcs (25x2)

Collection Tube 38 pcs 50 pcs (25x2)




well-1 Empty









well-10 Empty

Storage

MagPurix Forensic DNA Extraction Kit should be

stored at room temperature (15-25 °C). Do not

freeze the reagent cartridges. The Kits are stable

for 12 months under the condition





Starting Material

Whole blood, clotted/dried blood, Forensic Surface and Contact Swabs, hair roots,

saliva, sperm stain, chewing gum, cigarette butts, stamps, envelopes, tissue etc.

Sample preparation


Whole blood

(fresh or frozen)

To extract DNA from whole blood samples, please select and refer to MagPurix Blood DNA Extraction Kit 200 (ZP02001)

Clotted/ dried blood 1. Take 20μl blood sample to filter paper or bandage

2. Air-drying the blood sample

3. Cut the blood-contain range out, transfer pieces to

sample tube

4. Add 400μl BL2 and 20μl proteinase K to sample

5. Incubate at 56°C for 15min, vortex-mixing several

times during incubation or place sample in a

thermomixer

6. Transfer all the sample to filter column sitting in a

sample tube

7. Short spin at 500 x g, 1min

8. Take the sample tube for extraction

Forensic Surface and

Contact Swabs

1. Allow the swab or brush to air-dry for at least 2

hours after collection

2. Carefully cut or break off the end part of the swab

or brush into a 1.5ml microcentrifuge tube, using

an appropriate tool (e.g., scissors)

3. Add 200 or 400 μl of Buffer BL2 to the sample

4. Add 20 μl proteinase K, vortex-mixing for at least

10s

*If processing brush samples, centrifuge the tube

briefly (at 10,000 x g for 30 s) to force the brush to the

bottom of the tube

5. Incubate at 56°C for 15 min. vortex-mixing several



thermomixer


sample tube

7. Short spin at 500 xg, 1min


Hair root Using two or three 0.5–1 cm from the root ends of

plucked hair samples

【method1】

1. Place the hair sample in a 1.5 ml microcentrifuge

tube

2. Add 200μl Buffer BL2 to the sample.

3. Add 20μl proteinase K and 10 μl 1M DTT solution*,

and mix thoroughly by vortexing for at least 10 s

4. Incubate at 56°C for at least 6 h, vortex-mixing

several times during incubation or place sample in

a thermomixer

5. (optional) Add extra 10 μl proteinase K and 10 μl

DTT and incubate at 56°C until the hair samples are

completely dissolved

6. Spin the tube to remove drops from inside the lid


sample tube



*Prepare 1M DTT solution before processing the

protocol (1M is about 15% DTT(m/v))

【method2】

1. Place the hair sample in a 1.5 ml microcentrifuge

tube

2. Add 200 μl Buffer BL2 to the sample.

3. Add 20 μl proteinase K ,mix thoroughly by

vortexing for at least 10 s

4. Incubate at 56°C overnight, vortex-mixing several


thermomixer



sample tube




Human tissues Using up to 40mg tissue

1. Place tissue sample into a 1.5 ml microcentrifuge

tube

2. Add 200 or 400μl buffer BL2 and 20μl proteinaseK

to the sample, mix thoroughtly by vortexing for 10s

3. Incubate at 56°C for at least 2 hours,*

vortex-mixing several times during incubation or

place sample in a thermomixer

*Incubation for longer time (e.g. overnight) isn’t

making interferences of nucleic acid extraction



sample tube



Saliva 1. Place up to 50 μl saliva in a 1.5ml microcentrifuge

tube

2. Add 200 μl Buffer BL2 to the sample

3. Add 20 μl proteinase K, and mix thoroughly by

vortexing for 10 s

4. Incubate at 56°C for 15min, vortex-mixing several


thermomixer


6. Take 200μl to sample tube for extraction

Sperm stains 1. Place 5-10μl or 1cm2 of the forensic sample in a 1.5

ml centrifuge tube

2. Add 200 or 400μl Buffer BL2 to the sample


vortexing for 10 s

4. Incubate at 56°C for 15 min, vortex-mixing several


thermomixer

5. Spin the tube briefly to remove drops from inside

the lid



sample tube



Chewing gun Use of up to 40 mg of chewing gum cut into small

pieces is recommended

1. Place the chewing-gum sample in a 1.5ml


2. Add 200μl Buffer BL2 to the sample.


vortexing for 10 s.



thermomixer


the lid

6. Take 200μl sample to sample tube for extraction

Cigarette butts Use of approximately 1 cm2 paper from the end of the

cigarette or filter is recommended

1. Place the cigarette-butt sample in a 1.5 ml


2. Add 200 or 400μl Buffer BL2 to the sample

(Check if the sample has absorbed buffer BL2,if

necessary add more Buffer BL2 to the sample)


vortexing for 10 s



thermomixer


the lid


Stamps, envelopes Use of a 0.5–2.5 cm2 piece of postage stamp or

envelope is recommended

1. Place all the pieces of sample in a 1.5 ml


2. Add 200 or 400μ l Buffer BL2 to the sample

(Check if the sample has absorbed buffer BL2,if

necessary add more Buffer BL2 to the sample)



vortexing for 10 s



thermomixer


the lid






target








MagPurix Forensic DNA Extraction Kits is tested against predetermined specifications to



MagPurix Viral/Pathogen Nucleic Acids

Extraction Kit A

Cat. No. ZP02011 Process Time: 40-45 minutes (virus)

55-65 minutes (virus+bacteria)

Intended Use

MagPurix Viral/Pathogen Nucleic Acids Extraction Kit A is

used with the MagPurix 12 instrument for extraction of Viral

and bacterial DNA/RNA from cell-free samples, such as

serum, plasma, and other cell-free body fluids

Application

Nucleic acids extracted from MagPurix Viral/Pathogen

Nucleic Acids Extraction kit A can be used in a number of

downstream application including: PCR, qPCR,


Number Of Tests 48 extractions

Kit Components Kit Contents

(for 48 extractions)

Q’ty

Reagent Cartridge 48 pcs (4x6x2)

Reaction Chamber 48 pcs (4x6x2)

Tip Holder 48 pcs (24x2)

Filtered Tip 50 pcs (25x2)

Piercing Pin 50 pcs (25x2)

Sample Tube (2 mL) 50 pcs (25x2)

Elute Tube (1.5 mL) 50 pcs (25x2)

RNA Carrier (1mg) 1 pcs

Barcode Paper 1 pc












well-10 BL2 Buffer 400l

Storage

MagPurix Viral/Pathogen Nucleic Acids Extraction Kit A

should be stored at room temperature (15-25°C). Do

not freeze the reagent cartridges. The Kits are stable for

12 months under the condition

After dissolve the RNA carrier, store it at 4°C


not freeze–thaw the Frozen RNA carrier more than 3

times. Divide it into conveniently sized aliquots is

recommended


Starting Material



Elution Volume

Serum Total bacterial/

Viral Nucleic Acids

(DNA + RNA)

100-400μl(virus)

100-200(virus/bac

teria)

50-300μl Plasma

CSF

Pretreated Urine

Cell-free body

fluids

Controls/internal

control*


downstream analysis needed (*see control/internal control on page

76 )

This kit is designed for extraction of viral and bacterial

nucleic acids from plasma or serum, or from a pool of

such cell-free body fluids

After extraction, store the nucleic acid at 4°C (up to

24hours) or -20°C for longer storage. Repeated

freeze–thawing is not allowed

Sample

preparation

The purification procedure is optimized for use with 100-

400 μl serum, plasma*, CSF, pretreated urine or other

cell-free body fluid samples. ( *Blood samples treated

with EDTA or citrate as an anticoagulant can be used for

plasma preparation)

Samples can be either fresh or frozen, provided that they

have not been refrozen after thawing

RNA Carrier (CARRIER) serves two purposes during the

purification procedure. First, it enhances binding of viral

nucleic acids to the silica surface of the magnetic

particles, especially if the sample contains very few

target molecules. Second, the addition of large amounts

of RNA Carrier reduces the chances of RNA degradation


in the rare event that RNases are not denatured by the

chaotropic salts and detergent in the lysis buffer. If RNA

carrier is not added to the reaction, recovery of DNA or

RNA may be reduced

After collection and centrifugation, plasma, serum, or

CSF can be stored at 2–8°C for up to 6 hours. For longer

storage, we recommend freezing aliquots at –20°C

or –80°C. Thaw samples at room temperature (15–25°C),

and process the samples immediately when they have

equilibrated to room temperature. Do not refreeze the

aliquots after thawing. Repeated freeze–thawing leads to

denaturation and precipitation of proteins, resulting in

reduced viral titers and therefore reduced yields of

nucleic acids. If cryoprecipitates are visible in the

samples, centrifuge at 6800 x g for 3 minutes, transfer

the supernatants to fresh tubes without disturbing the

pellets, and start the purification procedure immediately

RNA Carrier Add 1.0 ml RNase-free water to lyophilized the RNA

Carrier (provided with the kit) and mix by vortexing

Store RNA Carrier at 4°C (short-term, up to 1 month) or

-20°C (long-term). Do not freeze–thaw the frozen carrier

RNA more than 3 times. Divide it into conveniently sized

aliquots is recommended

Before nucleic acid extraction, add RNA Carrier to the

sample is recommended. Add 5 μl Carrier RNA (for 100

μl sample), 10 μl (for 200 μl sample) or 20 μl (for 400 μl

sample) into the Sample Tube






target





Internal control(IC) Using a defined quantity control Place in sample tube or the round


Quality Control


MagPurix Viral/Pathogen Nucleic acids Extraction Kit is tested against predetermined

specifications to ensure consistent product quality.


MagPurix Viral/Pathogen Nucleic Acids

Extraction Kit B


Intended Use

MagPurix Viral/Pathogen Nucleic Acids Extraction Kit B is used with the MagPurix 12 instrument for extraction of viral and bacterial DNA/RNA from swab samples (cell-rich samples)

Application

Nucleic acids extracted from MagPurix Viral/Pathogen Nucleic Acids Extraction kit B can be used in a number of downstream application including: PCR, qPCR, Sequencing(NGS), Microarray, RFLP, Southern Blot Analysis

Number Of

Tests

48 extractions

Kit components

Kit Contents (for 48 extractions)

Q’ty








Barcode Paper 1pc












well-10 BL2 Buffer 400L

Storage

MagPurix Viral/Pathogen Nucleic acids Extraction Kit B

should be stored at room temperature (15-25°C). Do not

freeze the reagent cartridges. The Kits are stable for 12


Store the purified Nucleic acids at 4 °C (short- term) or

aliquot and store at -70°C (long-term) before perform the

downstream analysis


Starting

Material

Bacterial pellet/colony from culture, clinical swab samples

in liquid transport media, environment material (water, soil, etc.) and other cell-rich samples

Use the tissue or paraffin- embedded tissue sections(FFPE) as samples, we recommend to extract DNA by MagPurix Tissue DNA Extraction kit (ZP02004)


MagPurix Viral/Pathogen Nucleic Acids Extraction kit B purification procedures are shown in Table listed below,


Sample volume (Amount of starting material)

Elution Volume

Bacteria Pellet Total Viral/Bacterial Nucleic acids (DNA/RNA)

100-200 μl /Up to 109

bacteria (about OD600 =

3)

50-300 μl

Bacterial colony 100-200 μl /1-3 colony

Swab samples 100-200 μl liquid transport media

Controls/internal control*

Add controls /internal control in the extraction procedure if the downstream analysis needed (* see controls/internal control on page 82)

Sample

Preparation

Sample preparation requirements are highly dependent upon the type of starting material. Due to variations in consistency and viscosity, even similar sample types may require distinct handling

The table below describes the recommendations in processing the primary samples before nucleic acid extraction:


Table : Preparation of sample material for bacterial nucleic acid extraction


Inactivation the pathogenic microorganism

Recommended pretreatment : Boiling

4. Incubate samples at 95°C for 10 min

5. Centrifuge briefly to collect the complete

sample volume at the bottom of the tube

6. Allow samples to cool down or chill on ice, then

proceeding the following steps according to the

sample type

Viscous samples e.g. BAL, sputum or other mucous specimen

Recommended pretreatment : Liquefaction 1. Prepare a fresh DTT stock solution for

liquefaction * (e.g., 5× conc. DTT stock is about 0.75%)

2. Adjust the final DTT concentration in the sample

to 0.15% by adding DTT stock solution.

3. Incubate the sample (e.g., with shaking at 850r.p.m. for 30 min at 37°C) until it can be pipette easily.

4. Transfer 200 μL to sample tube (Supplied in the kit) * The liquefaction could be done by using other solutions, such as NALC（N-Acetyl-L-Cysteine）

-NaOH or other agents which could digest mucous material

For large volume liquid samples that have low or unknown bacterial loads e.g. urine, water collected from pool/river stream/tower

Recommended pretreatment : Centrifugation 1. Centrifuge the sample for up to 10 min at 20,000

× g to concentrate the bacterial cells in pellet

2. Discard supernatant, resuspend the pellet in 220 μL PBS

3. Take 200 μL to sample tube (Supplied in the kit)


Swab samples e.g. eye, nasal, pharyngeal, or other swabs

1. Collect samples and place in 1 ml PBS

containing a common fungicide. Incubate for 30min at room temperature

2. Take 200 μL to sample tube

For some gram-positive bacterial species. Especially for samples that contain particles e.g. stool

Recommended pretreatment : Mechanical homogenization

Follow the regular homogenization procedures in

the laboratory.

Bacterial suspension cultures

Take 200 μL culture to sample tube

Bacterial colony 1. Take 1-3 bacterial colony from culture plate with

an inoculation loop and suspend in 220 μL PBS by vigorous stirring






target







Quality Control


MagPurix Viral/Pathogen Nucleic acids Extraction Kit is tested against predetermined



MagPurix Viral RNA Extraction Kit


Intended Use

MagPurix Viral Nucleic RNA Extraction Kit is used with the

MagPurix 12 instrument for extraction of Viral RNA from

human biological specimens such as serum, plasma, and

other cell-free fluids

Application





Number Of Tests

48 extractions

Kit Components

Kit Contents ZP02013-48








RNA Carrier (1mg) 1pcs

Barcode Paper 1 pc












well-10 Empty

Storage

MagPurix Viral RNA Extraction Kit should be stored at


cartridges. The Kits are stable for 12 months under the

condition




times


Starting Material



Elution Volume

Serum Total Viral Nucleic

Acids (DNA + RNA)


CSF

Pretreated Urine

Cell-free body

fluids

Controls/internal

control#



86 )

The kit is designed for extraction of viral RNA from

plasma or serum, or from a pool of such cell-free body

fluids

After extraction, store the nucleic acid at 4°C (up to 1

hour) or -20°C for longer storage. Repeated


Sample

preparation


400 μl serum, plasma, CSF, or pretreated urine samples.

( Blood samples treated with EDTA or citrate as an

anticoagulant can be used for plasma preparation)













viralnucleic acids. If cryoprecipitates are visible in the




Carrier RNA For RNA virus, adding carrier RNA to the sample before

extraction is recommended !!!

Add 1.0 ml RNase free water to the carrier RNA tube

(provided with the kit) and mix by vortexing. Store it at

4°C (short-term, up to 1 month) or -20°C (long-term). Do


times

Add 5 μl Carrier RNA (for 100 μl sample), 10 μl (for 200 μl

sample) or 20 μl (for 400 μl sample) into the Sample

Tube





target








Quality Control


MagPurix Viral RNA Extraction Kits is tested against predetermined specifications to



MagPurix Plant DNA Extraction Kit


Intended Use

MagPurix Plant DNA Extraction Kit is used with the MagPurix 12 instrument for extraction of genomic DNA from plant (leaf, seeds and spores) and fungal tissues. Up to 100 mg of tissue can be used for purification

Application

Nucleic acids extracted from MagPurix Plant DNA Extraction




Number Of Tests

48 extractions

Kit Components





Filter Tip 50 pcs

Piercing Pin 50 pcs

Sample Tube (2 mL) 50 pcs

Elute Tube (1.5 mL) 50 pcs

Filter Column 50 pcs

Collection Tube 50 pcs

RNase A(10mg/mL) 1 pc (0.5mL)

Buffer PLA 1 pc (25mL)

Buffer PLB 1 pc (25mL)

Barcode Paper 1 pc



well-1 Empty









well-10 Empty

Storage

MagPurix Plant DNA Extraction Kit should be stored at



condition


store at -70°C (long-term) before perform the

downstream analysis


Starting Material After harvesting plant tissues, it should be frozen in liquid

nitrogen if not be used immediately. It can then be stored

at –80°C.Alternatively; tissue can be dried or lyophilized

after harvesting to allow storage at room temperature

(15–25°C). To ensure DNA quality, samples should be

completely dried within 24 hours of collection

If possible, it is preferable to collect young materials (e.g.,

leaves, needles) since they contain more cells per weight

and therefore result in higher yields

When working with fungi, harvest mycelium directly from

a culture dish or from liquid culture. For liquid culture,

first pellet cells by centrifugation. Remove the

supernatant completely before disruption and lysis.

Fresh, frozen, or freeze-dried fungal material can be use

The disruption method may require optimization to

ensure maximum DNA yield and quality. Complete and

quick disruption of starting material is essential to ensure

high DNA yields and to avoid DNA degradation

Before DNA extraction, plant material is first

mechanically disrupted with lysis buffer (Buffer PLA or

PLB). After Homogenization, remove the debris and other

precipitations by spin through the filter membrane.

Collect the clear flow-through and incubate with RNase A

to digest the RNA in the sample before DNA extraction

Sample preparation Using homogenizer to treat the tissue before extraction is

recommended

For better DNA yield, adding RNase A to sample before

extraction processing



Plant tissue 1. Add 440l lysis buffer* to sample

2. Perform homogenization by using

proper homogenizer

3. Remove Lysate to filter membrane

sitting in collection tube

4. Short spin at 6000 xg to collect clear

flow-through

5. Add 10l RNase A, incubate for 10min

at room temperature

6. Transfer to sample tube

7. Perform extraction

Yeast A. Suspension culture

1. Centrifuge at 6000xg, 3min

2. Remove supernatant

3. Add 440mL plant lysis buffer*,

vortex mixing 30 sec

4. Transfer 400 L to sample tube


B. Culture colony

3. Take 1-3 colony from culture plate with an inoculation loop and suspend in 440 μL of plant lysis buffer* by vigorous stirring



*We provide two kind plant lysis buffer: PLA and PLB for

dealing with different tissue types. Before extraction a new

tissue type, try these two lysis buffer for getting the optimized

lysis procedure and a better DNA yield






target








MagPurix Plant DNA Extraction Kits is tested against predetermined specifications to



MagPurix Total RNA Extraction Kit


Intended Use

MagPurix Total RNA Extraction Kit is used with the MagPurix

12 instrument for extraction of total RNA from whole blood,

blood cells, animal tissue, plant tissue, yeast or cultured cells

Application

Total RNA extracted from MagPurix Total RNA Extraction


including: RT-PCR, qPCR, Sequencing(NGS), Microarray,

Northern blotting

Number Of Tests 36 and 48 extractions

Kit Components Kit Contents Quantity

ZP02015-36 ZP02015-48



Tip Holder 36 pcs (6X6) 48 pcs (6X4x2)

Filter Tip 38 pcs 50 pcs

Piercing Pin 38 pcs 50 pcs

Sample Tube (2 mL) 76 pcs (38X2) 100 pcs (50X2)

Elute Tube (1.5 mL) 38 pcs 50 pcs

RL A Buffer 25ml 25ml

RL B Buffer 25ml 25ml

Filter column 38 pcs 50 pcs

Collection tube 38 pcs 50 pcs













well-10 DNase buffer 1000 μl

Storage

MagPurix Total RNA Extraction Kit should be stored at



condition

After extraction, store RNA at -60 to -80℃

immediately, repeated freeze–thawing is not allowed.

Always handle RNA on ice for downstream analysis


Purification Protocol

Protocol name Sample vol.

Elute vol.

Description

Total RNA 100-400l

50-200l

Extraction Total RNA and DNA from

sample

Total RNA

(DNA-free)

100-400l

50-200l

Extraction total RNA (DNA-free)* from

sample

* Before use protocol-Total RNA (DNA-free), prepare DNase (not supplied in the kit)

is necessary

Before starting

-Mercaptoethanol (β-ME) must be added to Buffer RLT before use

If performing DNA-free protocol, Prepare DNase before extraction. Place 10 l

DNase in sample tube( at the middle well of sample rack) when extraction

Homogenization is necessary for animal tissue, Plant tissue and yeast before

extraction. Add RL Buffers to sample when perform homogenizing

Wear clean glove, use RNase-free filter tip, and keep work area, pipettors and

reagents free of virus, bacteria and Nuclease contamination

Using RNaseZap® to clean the surface of bench, equipments and pipettors is

one of the most easy way to remove the RNase contaminations of work area

Using RNA stabilized reagent (e.g. RNA later) to treat sample is one of the best

way to protect the RNA if the sample cannot be processing in a RNase-free

working area


Reagents to be supplied by user

Reagent Description Preparation β -mercaptoethanol

(β -ME)

β -ME reduce disulfide

bonds and irreversibly

denature the RNase and

eliminate RNase released

during cell lysis

Add 10 μl β-ME per 1 ml RL

lysis Buffers*. It can be

stored at RT for up to one

month

Red blood cells lysis

buffer

(RBC lysis buffer)

Lyse Erythrocyte from

whole blood

(Erythrocyte (RBC) lysis

procedure)

10xRBC lysis buffer(100ml)

8.29g NH4Cl (1.5M)

1g KHCO3 (100mM)

0.0372g Na2EDTA (10mM)

Adjust pH7.2-7.4 by HCl

0.2 mm filtered, store for 6

months at 4℃

Dilute 10 times fresh before

use

DNase

To liminate DNA

contamination

Novagen RNase-free DNaseI

(69182-3CN)

*RL lysis buffers means RLA and RLB Buffers. Dispense the β-ME in a fume hood and

wear appropriate protective clothing

Starting Material

Whole Blood

1. Using the fresh whole blood sample for isolation. (within 4 hr, on ice) Freezing

blood is not allowed. The blood sample should be collected in the presence of

an anticoagulant, preferably EDTA, although other anticoagulants such as

citrate, heparin, or ACD (acid citrate dextrose) can also be used.

2. For optimal results, blood samples should be processed within a few hours of

collection and keep in 4℃.

3. Performing Erythrocyte (RBC) lysis procedure before extraction.

4. Using the whole blood samples which have extreme high WBCs no. (more than

10000/l) or concentrated PBMCs( peripheral blood mononucleated cells),

decrease the input volume for extraction is recommended (total WBC no. less

than 5 x 106).


Tissue

1. To prevent degradation by intracellular RNase, it is important that tissues are

either flash-frozen in liquid nitrogen or stored at –70°C, or processed

immediately following excision.

2. Using RNA stabilized reagent (e.g. RNA later) to treat tissue is another option to

protect the RNA if the sample cannot be froze immediately. Frozen tissue

should not be allowed to thaw during handling (e.g., weighing), keep sample on

ice during cutting or homogenized with RLA Buffer is recommended.

3. After homogenization, using filter column (supplied in the kit) to remove the

insoluble and viscous material of the lysates.

Cells

1. Cells or isolated blood cells can be collected as pellets and either flash-frozen in

liquid nitrogen and stored at –70°C, or processed immediately. Add RLA Buffer

to resuspend pellet for extraction

2. Alternatively, samples can be stored at –70°C in RLA Buffer after disruption and

homogenization. Samples frozen in this way are stable for months.

Plant tissue and yeast

1. Up to 100 mg of sample is first ground in liquid nitrogen or frozen, then add

lysis buffer(RLA or RLB Buffer) to homogenizer

2. Most Plant cells use RLA Buffer for disruption and denaturing sample

3. However, some tissues, such as milky endosperm of maize or mycelia of

filamentous fungi, solidify in RLA Buffer, making the extraction of RNA

impossible. In these cases, RLB Buffer should be used instead.

4. After add lysis buffer(RLA or RLB), samples are place into homogenizer for

homogenization

5. After homogenization, using filter column (supplied in the kit) to remove the

insoluble and viscous material of the lysates.

It is essential to use the correct amount of starting material in order to obtain

optimal RNA yield and purity (as the table below). Use excess quantity is not

helpful in total RNA extraction.


Sample Type Sample Volume (Amount of starting material)

Elution Volume

Whole blood 200-400 l*

(WBCs no. about 106)

50-200μl**

PBMCs Up to 50 l

(suspended in 200l with RL buffer)

Tissue 10-40mg

(Lysed and suspend with RL buffer)

Cultured cells 200-400 l suspension of primary or

cultured cells (cell no. < 5 x 106)

Plant tissue Up to 100mg

Yeast Up to 100mg

Controls/internal

control


downstream analysis needed

* Blood cells needs to perform manual RBC lysis procedure

before extraction

** After extraction, store RNA at -60 to -80℃ immediately,

repeated freeze–thawing is not allowed

Sample preparation

Sample Procedure

Whole blood 1. Fresh prepare 1x RBC lysis buffer

2. Add ice-cold two volume RBC lysis buffer to one

volume blood sample

3. Inverting 3-5 times, incubate on ice for 10-15 min

4. Centrifuge at 1000 x g, 10min, 4℃


6. Resuspend pellet with 220 l RLA Buffer

7. Take 200 l for extraction

PBMCs

(Peripheral Blood

Mononucleated Cells)

1. Resuspend PBMCs with 220l RLA Buffer

2. Vortex mixing for 10 sec

3. Take 200 l for extraction


Tissue 1. Add 220l RLA Buffer to tissue; make sure the

sample if completely immersed in buffer. Increase

RLA buffer input amount up to 440l if tissue

sample is large.

2. Homogenized tissue by homogenizer

3. Spin down the lysate

4. Remove all the lysate to filter column sitting in

collection tube

5. Centrifuge at 1000 x g , for 5min on 4℃

6. Transfer 200-400l to sample tube


Cultured cells 【Protocol 1】Suspension culture

1. Harvest cell culture

2. Centrifuge at 1000xg, 5min on 4℃

3. Remove supernatant completely

4. Resuspend cell pellet with 22oml RLA Buffer

5. Vortex mixing for 10 sec

6. Take 200l for extraction

【Protocol 2-1】Monolayer culture

1. Trypsinize the cells

2. Harvest the cell in PBS

3. centrifuge at 300xg, 5min on 4℃


5. Resuspend pellet with 220 l RLA Buffer

6. Vortex mixing for 10sec

7. Take 200l for extraction

【Protocol 2-2】Monolayer culture

1. Scrape the cells with 220-440l RLA Buffer

2. Vortex mixing for 10sec

3. Take 200-400l for extraction

Plant tissue/ Yeast 1. Add 220 -440l RLA or PLB Buffer* to sample, make

sure the sample if complete immersed in buffer

2. Homogenized tissue by homogenizer

3. Remove lysate to filter column sitting in collection

tube


4. Centrifuge at 1000 x g , for 5min on 4℃

5. Spin down

6. Transfer 200-400l to sample tube


DNA-free RNA

extraction

1. After sample preparation

2. Remove sample to sample tube at top well of

sample rack

3. Place 10 ml DNase in sample tube at the middle well

of sample rack

4. Place elute tube at bottom well of sample rack

5. Select “Total RNA (DNA-free)” protocol to start

extraction





target





Internal control(IC) Using a defined quantity control Place in the round well of the reaction

chamber

Quality Control


MagPurix Total RNA Extraction Kit is tested against predetermined specifications to



MagPurix Viral Nucleic Acid Extraction Kit 800


Intended Use

MagPurix Viral Nucleic Acid Extraction Kit 800 is used with

the MagPurix 12 instrument for extraction of Viral DNA or

RNA from human biological specimens such as serum,

plasma, and other cell-free fluids.

Application



applications including: PCR, qPCR, Sequencing(NGS),


Number Of Tests

48 extractions

Kit Components





Filter Tip 50 pcs

Piercing Pin 50 pcs

Sample Tube (2 mL) 50 pcs

Elute Tube (1.5 mL) 50 pcs

RNA Carrier (1mg) 1 pcs

Barcode Paper 1 pc












well-10 Empty

Storage

MagPurix Viral Nucleic Acid Extraction Kit 800 should be

stored at room temperature (15-25°C). Do not freeze






times


Starting Material



Elution Volume

Serum Total Viral Nucleic

Acids (DNA + RNA)


CSF

Pretreated Urine

Cell-free body

fluids

Controls/internal

control#



24 )

The kit is designed for extraction of viral nucleic acids

(e.g., those of HIV, HCV, HBV, or HPV) from plasma or

serum, or from a pool of such cell-free body fluids

After extraction, store the nucleic acid at 4°C (up to

24hours) or -20°C for longer storage. Repeated


Sample

preparation


400 μl serum, plasma, CSF, or pretreated urine samples.

( Blood samples treated with EDTA or citrate as an

anticoagulant can be used for plasma preparation)













viralnucleic acids. If cryoprecipitates are visible in the




Carrier RNA For RNA virus, adding carrier RNA to the sample before

extraction is recommended !!!

Add 1.0 ml RNase free water to the carrier RNA tube

(provided with the kit) and mix by vortexing. Store it at

4°C (short-term, up to 1 month) or -20°C (long-term). Do


times

Add 5 μl Carrier RNA (for 100 μl sample), 10 μl (for 200 μl

sample) or 20 μl (for 400 μl sample) into the Sample

Tube





target





Internal control(IC) Using a defined quantity control Place in sample tube or the round well of

the reaction chamber*

*Use RNA as internal control, add it into round well of reaction chamber is recommended


MagPurix Viral Nucleic Acid Extraction Kits 800 is tested against predetermined



Protocol of extraction

1. Turn the power switch on and waiting for the LCM

screen turn on and shows “MagPurix 12 System

Stand-By”.

2. Press “Start” button

(The system will process self-testing, and then go to

steady mode)

Note:

The system will block main functions before the

completion of self-testing process.

3. Open the sliding door and remove the sample rack

from the instrument.

4. Load Reagent Cartridges, and all plastics disposables

(Reaction Chamber, Tip Holder, Piercing Pin, Filtered

Tip and Pestle (optionally supplied with some kit types)

Insert the cartridges


How to pull apart reagent cartridges

Slash open the dotted line with nail and snap it with a little bit

force.

Insert Reaction Chambers


Insert Tip Holder

Insert Piercing Pins

Insert Filtered Tips


Note:

1. The positions of piercing pin and filtered tip; the 2nd

one should be “EMPTY”.

2. Load one Reagent Cartridge and one set of plastic

disposable per sample.

Important:

- Set Cartridges in the order of the number from

left to right.

- Make sure that Cartridges are inserted in to the

Cartridge Tray tightly.

- You can load 1-12 cartridges on the tray

depending on the number of samples that you

wish to process.

5. Load Sample Tube and Elute Tube to Sample Rack on the

bench


Insert Sample Tube on to Sample Rack

Insert Elute Tube on to Sample Rack

6. Load the sample(s) to Sample Tube

Note:

- Pretreatments are essential for some sample

types before loading to Sample Tube. Please refer

to the handbook of reagent kits for details.

- Make sure the caps of Elute Tube are open as the

figure shown above.


7. Place Sample Rack on the instrument platform

Note:

- Use two hands to handle the Sample Tray.

- Make sure the Sample Tray be placed correctly on to

the instrument

8. Close the door

9. Scan the protocol barcodes to select purification protocol,

sample volume and elute volume

Note:

- There is one protocol barcode paper enclosed in

the reagent kit box

- Protocol’s name, sample volume and elution

volume will be shown on LCM screen after

protocol barcode is scanned

10. Follow the instructions displayed on LCM screen to double

check the operating steps being completed before

program running.


11. Push “Enter” to confirm. Instrument will start to run the

protocol program automatically until whole processes are

completed.

Note:

- It takes from 30 to 45 minutes to complete the

extraction according to reagent types

12. At the end of the run, the instrument beeps briefly and the

LCM shows “Protocol Completed”

13. Open the instrument door

14. Remove the elute tubes containing the purified nucleic

acid

Note: Store the purified nucleic acids at 4℃ for

short-term storage or store at -70℃ for long-term

storage

15. Discard the used cartridges, all plastic consumables into

biohazard waste. Do not reuse the cartridges

16. If you’re not using the instrument, place the Sample Rack

back to workplace, close the instrument door and push

“Start” button for 2 secs to get into “sleeping mode”.

And for longer time not using the instrument turn the

power switch off.


Zinexts Life Science Corporation

www.zinexts.com

2F.-2, No.122, Qiaohe Rd., Zhonghe Dist.,

New Taipei City 235, Taiwan

Tel: +886 2 2246 3579

Fax: +886 2 2243 8570

Mail: [email protected]

http://www.zinexts.com/

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