Endpoint PCR Real-Time PCR Reverse Transcription · Lyophilisation (= freeze-drying) is a...

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Molecular Biology › › Endpoint PCR › › Real-Time PCR › › Reverse Transcription › › dNTPs › › Custom-Specific Lyophilisation Lyophilisation

Transcript of Endpoint PCR Real-Time PCR Reverse Transcription · Lyophilisation (= freeze-drying) is a...

Page 1: Endpoint PCR Real-Time PCR Reverse Transcription · Lyophilisation (= freeze-drying) is a dehydra-tion method applying low temperatures and reduced ambient pressure. In these conditions

Molecular Biology

ACCCACGAAAGGGAA ATAAGC AACO TTCAGGGAAGAA CTAUAACTGCCAC ACCCACGAAAGGGAA ATAAGC AACO TTCAGGGAAGAA CTAUAACTGCCAC ACCCACGAAAGGGAA ATAAGC AACO TTCAGGGAAGAA CTAUAACTGCCAC ACCCACGAAAGGGAA ATAAGC AACO TTCAGGGAAGAA CTAUAACTGCCAC ACCCACGAAAGGGAA ATAAGC AACO TTCAGGGAAGAA CTAUAACTGCCAC ACCCAC-GAAAGGGAA ATAAGC AACO TTCAGGGAAGAA CTAUAACTGCCAC ACCCACGAAAGGGAA ATAAGC AACO TTCAGGGAAGAA CTAUAACTGCCAC ACCCACGAAAGGGAA ATAAGC AACO

›› Endpoint PCR›› Real-Time PCR›› Reverse Transcription›› dNTPs›› Custom-Specific Lyophilisation

Lyophilisation

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2 Introduction

Introduction

No fridge

required.

Building Blocks of Life

›› ready-to-use›› individual scales›› customized packaging

& labeling

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Nucleotides & Nucleosides

In our chemistry division, we have hundreds of natural and modified nucleotides in stock. In addition, with our pre-made building blocks and in-house expertise we manufac-ture even the most exotic nucleotide analog from mg to kg scale.

LEXSY Expression

In the field of recombinant protein production, Jena Bioscience has devel-oped its proprietary LEXSY (Leishmania Expression System) technology. It is based on an S1-classified unicellular organism that combines easy handling with a eukaryotic protein folding and modification machinery. Besides eve-rything you need to establish LEXSY in your lab we also offer custom expres-sion of recombinant proteins.

Molecular Biology & ProteinsFor applications in the field of Molecular Biology we offer a large selection of single reagents, complete kits and optimized master mixes. This section includes products for DNA or RNA purification, amplification and modification with focus on PCR-related techniques.

For your questions regarding lyophilisation contact me directly: [email protected]

Click Chemistry, Probes & Epigenetics

Our Probes & Epigenetics as well as Click Chemistry sections offer inno-vative reagents for the functionaliza-tion, conjugation and labeling (fluo-rophores, haptens) of (bio) molecules complemented by epigenetic modifi-cation analysis tools.

Crystallography & Cryo-EM

For the crystallization of biological macro-molecules – which is often the bottleneck in determining the 3D-structure of proteins – we offer specialized reagents for protein sta-bilization, crystal screening, crystal optimization, and phasing that can reduce the time necessary to obtain a high resolution protein structure from several years to a few days.

3

Building Blocks of Life

www.jenabioscience.com Introduction

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4 Building Blocks of Life

Imprint: Design & Layout by timespin - Digital Communication GmbH, Sophienstr. 1, D-07743 Jena, Germany, www.timespin.de Copyright: Please contact Jena Bioscience if you want to use texts and/or images in any format or media.

Established in 1998 by a team of scientists from the Max-Planck-Institute of Molecular Physiology (Dortmund), Jena Bioscience utilizes more than 25 years of academic know-how to develop innova-tive reagents for clients from both research and industry in 100+ countries. To date, Jena Bioscience still remains an owner-operated business.

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5www.jenabioscience.com IntroductionACCCACGAAAGGGAA ATAAGC AACO TTCAGGGAAGAA CTAUAACTGCCAC ACCCACGAAAGGGAA ATAAGC AACO TTCAGGGAAGAA CTAUAACTGCCAC ACCCACGAAAGGGAA ATAAGC AACO TTCAGGGAAGAA CTAUAACTGCCAC ACCCACGAAAGGGAA ATAAGC AACO TTCAGGGAAGAA CTAUAACTGCCAC ACCCACGAAAGGGAA ATAAGC AACO TTCAGGGAAGAA CTAUAACTGCCAC ACCCACGAAAGGGAA ATAAGC AACO TTCAGGGAAGAA CTAUAACTGCCAC ACCCACGAAAGGGAA ATAAGC AACO TTCAGGGAAGAA CTAUAACTGCCAC ACCCACGAAAGGGAA ATAAGC AACO

Contents

Introduction .............................................6

Endpoint PCR ........................................... 7

Real-Time PCR ........................................8

Reverse Transcription ............................9

dNTPs .......................................................10

Custom-Specific Lyophilisates .......... 11

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6 Building Blocks of LifeIntroduction

Lyophilisation (= freeze-drying) is a dehydra-tion method applying low temperatures and reduced ambient pressure. In these conditions water sublimes from a solid state directly into the gas phase (Fig. 1). As a result, the moisture content of a product is decreased to approxi-mately 1 %[1]. In the absence of water growth of microorganisms and chemical (degradation) reactions are inhibited leading to long-term preservation. Freeze-dried materials may be stored at ambient temperatures for many years while remaining structurally & chemically sta-ble. They are fully reconstituted after addition of water (rehydration).

Introduction

Reference:[1] Patel et al (2011) Emerging freeze-drying process development and scale-up issues. Aaps Pharmscitech 12 (1): 372-378.

When to consider freeze-drying?

Modern freeze-drying techniques revolutionized preservation of perishable materials not only for chemi-cal or food industries. Lyophilisation also provides significant advantages to many of todays molecular biological applications and products:

Figure 2 Jena Bioscience in-house data show that freeze-dried PCR mixes remain fully functionals after 12 month heat exposure (30 °C, green circle) while liquid mixes significantly lose activity (4 °C, red circle).(Assay: Amplification of a 4 kb fragment, lambda DNA, template dilution series)

1. Due to the mild operating conditions (low pressure & temperatures; Fig. 1) freeze drying is particularly suitable for (heat-) sensitive products (e.g. enzymes)

2. Lyophilisation dramatically enhances shelf life, which is especially useful for products with poor stability in aqueous solutions (e.g. proteins)

3. Since dry mixes require no refrigeration, they are stored and handled at room temperature. At the same time, no activity loss caused by freeze-thaw cycles occurs.

Figure 1 Phase Diagram of Water

Lyophilisate Liquid Mix

Good to know

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7www.jenabioscience.com Endpoint PCR

Jena Bioscience lyophilisates are ready-to-use and preloaded in a dry, room temperature stable format.

Mixes are available in preferred packaging such as:

§ single PCR reaction tubes § 8-tube strips (low/high profile) § 96-well plates (low/high profile; flat top,

half/full skirt) § custom-specific formats

Cat.-No. Amount Price (EUR)PCR-160S 192 reactions x 20 µl 200,00PCR-160L 960 reactions x 20 µl 800,00

Direct PCR Lyophilisates are designed for amplification of target DNA directly from whole blood, ani-mal and plant tissues. No pretreatment or prior purification of DNA is required.

Endpoint PCR

Multiplex PCR Master Lyophilisate is especially designed for parallel amplification of a variety of frag-ments in a single PCR assay. The mix is suitable for multiple target gene amplification in a single tube.

Cat.-No. Amount Price (EUR)PCR-161S 192 reactions x 20 µl 212,50PCR-161L 960 reactions x 20 µl 850,00

Crystal Taq Master Lyophilisate recommended for routine PCR applications (up to 4 kb fragment length), high throughput PCR or genotyping.

Cat.-No. Amount Price (EUR)PCR-152S 192 reactions x 20 μl 93,90 PCR-152L 960 reactions x 20 μl 375,60

Crystal Hot Start Master Lyophilisate recommended when amplifying low-copy-number targets, working with complex backgrounds or when prolonged room-temperature set-ups are unavoidable.

Cat.-No. Amount Price (EUR)PCR-153S 192 reactions x 20 μl 114,70 PCR-153L 960 reactions x 20 μl 458,80

Preloaded and stable at Room Temperature

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8 Building Blocks of LifeReal-Time PCR

JBS qPCR ProbesMaster Lyophilisate is based on detection of labeled DNA probes. Mixes are recom-mended for use with Dual Labeled Fluorescent Probes, e.g. TaqMan®, Molecular Beacons or FRET probes.

Cat.-No. Amount Price (EUR)

qPCR ProbesMaster LyophilisatePCR-156S 192 reactions x 20 μl 137,50 PCR-156L 960 reactions x 20 μl 550,00

qPCR ProbesMaster Lyophilisate highROXPCR-168S 192 reactions x 20 µl 137,50PCR-168L 960 reactions x 20 µl 550,00

qPCR GreenMaster Lyophilisate is based on fluorescent DNA stain fluorescent DNA stain EvaGreen™. No sequence-specific labeled probes are needed.

qPCR GreenMaster LyophilisatePCR-157S 192 reactions x 20 μl 137,50 PCR-157L 960 reactions x 20 μl 550,00

qPCR GreenMaster Lyophilisate highROX PCR-170S 192 reactions x 20 µl 137,50 PCR-170L 960 reactions x 20 µl 550,00

qPCR SybrMaster Lyophilisate is based on fluorescent DNA fluorescent DNA stain SYBR® Green. No sequence-specific labeled probes are needed.

qPCR GreenMaster LyophilisatePCR-173S 192 reactions x 20 µl 137,50 PCR-173L 960 reactions x 20 µl 550,00

qPCR SybrMaster Lyophilisate highROXPCR-174S 192 reactions x 20 µl 137,50 PCR-174L 960 reactions x 20 µl 550,00

Real-Time PCR

Jena Bioscience Real-Time PCR lyophilisates are designed for quantitative DNA analysis in real-time based on flourescent DNA stains or labeled probes.

All mixes are also available in combination with high ROX as reference dye for cycler-internal signal normalisation.

No freeze-thaw cycles – No more pipetting on ice.

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9 Reverse TranscriptionACCCACGAAAGGGAA ATAAGC AACO TTCAGGGAAGAA CTAUAACTGCCAC ACCCACGAAAGGGAA ATAAGC AACO TTCAGGGAAGAA CTAUAACTGCCAC ACCCACGAAAGGGAA ATAAGC AACO TTCAGGGAAGAA CTAUAACTGCCAC ACCCACGAAAGGGAA ATAAGC AACO TTCAGGGAAGAA CTAUAACTGCCAC ACCCACGAAAGGGAA ATAAGC AACO TTCAGGGAAGAA CTAUAACTGCCAC ACCCACGAAAGGGAA ATAAGC AACO TTCAGGGAAGAA CTAUAACTGCCAC ACCCACGAAAGGGAA ATAAGC AACO TTCAGGGAAGAA CTAUAACTGCCAC ACCCACGAAAGGGAA ATAAGC AACO

Reverse Transcription

One-Step RT-PCR Master Lyophilisate is designed for performing maximum sensitive and specific RT-PCRs convenient in single tubes. The enzyme mix is based on a genetically engineered RT poly-merase with enhanced thermal stability resulting in an increased specificity, higher cDNA yield and an improved efficiency for highly structured and long cDNA fragments.

Cat.-No. Amount Price (EUR)PCR-159S 192 reactions x 20 μl 475,00 PCR-159L 960 reactions x 20 μl 1.900,00

RT Master Lyophilisate contains a genetically engineered version of M-MLV Reverse Transcriptase (M-MLV RT) with eliminated RNase H activity and increased thermal stability. The enzyme is a RNA-directed DNA polymerase that synthesizes a complementary DNA strand initiating from a primer using single-stranded RNA or DNA as template.

Cat.-No. Amount Price (EUR)PCR-158S 192 reactions x 20 μl 370,00 PCR-158L 960 reactions x 20 μl 1.480,00

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10 Building Blocks of LifedNTPs

dNTPsJena Bioscience's enzymatic dNTP manufacturing process ensures superior quality for deoxynucleo-tide triphosphates: All our sodium salt dNTP lyophilisates are ultrapure (>95% by HPLC) and function-ally tested by a set very sensitive enzymatic reactions.

Cat.-No. Amount Price (EUR)

dATP - SolidNU-1043-20 20 μmol 20,46 NU-1043-100 5 x 20 μmol 81,84

dCTP - SolidNU-1044-20 20 μmol 20,46 NU-1044-100 5 x 20 μmol 81,84

dGTP - SolidNU-1045-20 20 μmol 20,46 NU-1045-100 5 x 20 μmol 81,84

dTTP - SolidNU-1046-20 20 μmol 20,46 NU-1046-100 5 x 20 μmol 81,84

dUTP - SolidNU-1041-20 20 μmol 24,55NU-1041-100 5 x 20 μmol 98,21

dITP - SolidNU-1040-20 20 μmol 24,55NU-1040-100 5 x 20 μmol 98,21

EquipmentThere is a great diversity of freeze-dryers on the market. At the most basic level however, they all share 4 essential components:

§ Chamber (to hold samples) § Refrigerating system (to cool down samples) § Vacuum pump (to reduce chamber pressure) § Condenser (to remove sublimed water)

Jena Biosciences uses a compact two-chamber freeze-dryer from Zirbus Technology (Figure 3). The microprocessor-controlled system enables flexible adjustment of desired process parame-ters (e.g. shelf temperature, chamber pressure) and continuous documentation via its internal Zirbus Dry Control operating and recording tool.

Figure 3 Foto by courtesy of Christian Zirbus, Zirbus technology GmbH

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11www.jenabioscience.com Custom-Specific Lyophilisates

Custom-Specific LyophilisatesLooking for freeze-dried mixes beyond the standard Jena Bioscience repertoire?But … … you are reluctant to take the high costs of equipment acquisition?

… you need a reliable partner with the necessary technical advice and know how?

We manufacture individualized lyophilisates according to your requirements (including addition of primers or DNA controls) and applications (Endpoint PCR, Real-Time PCR, Reverse Transcription).

Based on your liquid mixes we develop formulations, lyophilise at desired scales and offer customized product packaging & labeling.

Liquid Mix

Scale-Up QualityControl

Individual freeze-dried mix

Custom - Dispensing- Packaging- Labeling

Pilot-Scale Optimization

1.Formulation

Development

2. Small-scale testing

- Functionality- Stability- Appearance

3. Re-formulation- Stabilizers- Protectors- Process parameters

Manufactured under ISO 9001 reagulations and validated by customer on-site audits Jena Bioscience guarantees long-term product stability (≥ 6 months at 30 °C).

E-Mail me for service inquiries or further information: [email protected]

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Jena Bioscience GmbHLoebstedter Str. 7107749 JenaGermany

Phone +49 (0)3641- 62 85 000Fax +49 (0)3641- 62 85 [email protected]

www.jenabioscience.com

Molecular Biology

ACCCACGAAAGGGAA ATAAGC AACO TTCAGGGAAGAA CTAUAACTGCCAC ACCCACGAAAGGGAA ATAAGC AACO TTCAGGGAAGAA CTAUAACTGCCAC ACCCACGAAAGGGAA ATAAGC AACO TTCAGGGAAGAA CTAUAACTGCCAC ACCCACGAAAGGGAA ATAAGC AACO TTCAGGGAAGAA CTAUAACTGCCAC ACCCACGAAAGGGAA ATAAGC AACO TTCAGGGAAGAA CTAUAACTGCCAC ACCCAC-GAAAGGGAA ATAAGC AACO TTCAGGGAAGAA CTAUAACTGCCAC ACCCACGAAAGGGAA ATAAGC AACO TTCAGGGAAGAA CTAUAACTGCCAC ACCCACGAAAGGGAA ATAAGC AACO

Request a sample!

Test the exceptional performance of our lyophilisates for yourself.

Our entire product range is available as samples.


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