Molecular marker technology in studies on plant genetic diversity
-
Upload
chanakya-pachi -
Category
Education
-
view
343 -
download
6
Transcript of Molecular marker technology in studies on plant genetic diversity
![Page 1: Molecular marker technology in studies on plant genetic diversity](https://reader036.fdocuments.net/reader036/viewer/2022062412/58838cd41a28ab07628b69db/html5/thumbnails/1.jpg)
Molecular marker technology in studies on plant genetic diversity
PCR based techniques
PCR with arbitrary primers
PCR with Amplified fragment
length polymorp
hisms
Sequence-tagged sites
Microsatellites,
SCARs, CAPS, ISSRsBy
Chanakya Pachi
![Page 2: Molecular marker technology in studies on plant genetic diversity](https://reader036.fdocuments.net/reader036/viewer/2022062412/58838cd41a28ab07628b69db/html5/thumbnails/2.jpg)
Molecular Markers
• The development of molecular techniques for genetic analysis has led to a great augmentation in our knowledge of crop genetics and our understanding of the structure and behavior of various crop genomes. • These molecular techniques, in particular the
applications of molecular markers, have been used to scrutinize DNA sequence and it’s variation(s) in and among the crop species and create new sources of genetic variation by introducing new and favorable traits from landraces and related crop species.
![Page 3: Molecular marker technology in studies on plant genetic diversity](https://reader036.fdocuments.net/reader036/viewer/2022062412/58838cd41a28ab07628b69db/html5/thumbnails/3.jpg)
Markers can aid selection for target alleles that• Are not easily
assayed in individual plants,
• Minimize linkage drag around the target gene, and
• Reduce the number of generations required to recover a very high percentage of the recurrent parent genetic background
![Page 4: Molecular marker technology in studies on plant genetic diversity](https://reader036.fdocuments.net/reader036/viewer/2022062412/58838cd41a28ab07628b69db/html5/thumbnails/4.jpg)
Introduction
• A genetic marker is a gene or DNA sequence with a known location on a chromosome and associated with a particular gene or trait. • It can be described as a variation, which may arise
due to mutation or alteration in the genomic loci, that can be observed. • A genetic marker may be a short DNA sequence,
such as a sequence surrounding a single base-pair change (single nucleotide polymorphism, SNP), or a long one, like minisatellites.
![Page 5: Molecular marker technology in studies on plant genetic diversity](https://reader036.fdocuments.net/reader036/viewer/2022062412/58838cd41a28ab07628b69db/html5/thumbnails/5.jpg)
Some commonl
y used types of genetic markers
RFLP (or Restriction fragment
length polymorphis
m)
AFLP (or Amplified fragment
length polymorphis
m
RAPD (or Random
amplification of
polymorphic DNA)
VNTR (or Variable number tandem repeat)
Microsatellite polymorphis
m SNP (or Single
nucleotide polymorphis
m)
STR (or Short tandem
repeat)
SFP (or Single feature
polymorphism)
DArT (or Diversity Arrays
Technology)
![Page 6: Molecular marker technology in studies on plant genetic diversity](https://reader036.fdocuments.net/reader036/viewer/2022062412/58838cd41a28ab07628b69db/html5/thumbnails/6.jpg)
DNA-based technologies•PCR based techniques • PCR with arbitrary primers• Amplified fragment length polymorphisms (AFLPs)• Sequence-tagged sites• Microsatellites• SCARs• CAPS• ISSRs
![Page 7: Molecular marker technology in studies on plant genetic diversity](https://reader036.fdocuments.net/reader036/viewer/2022062412/58838cd41a28ab07628b69db/html5/thumbnails/7.jpg)
PCR BASICS• The polymerase chain reaction• PCR is a rapid, inexpensive and simple
way of copying specific DNA fragments from minute quantities of source DNA material • It does not necessarily require the use of
radioisotopes or toxic chemicals• It involves preparation the sample DNA • A master mix with primers, • Followed by detecting reaction products
![Page 8: Molecular marker technology in studies on plant genetic diversity](https://reader036.fdocuments.net/reader036/viewer/2022062412/58838cd41a28ab07628b69db/html5/thumbnails/8.jpg)
PCR procedures: steps• Denaturation: • DNA fragments are heated at high
temperatures, which reduce the DNA double helix to single strands. • These strands become accessible to
primers• Annealing: • The reaction mixture is cooled down.• Primers anneal to the complementary
regions in the DNA template strands, and double strands are formed again between primers and complementary sequences
• Extension: • The DNA polymerase synthesises a
complementary strand. • The enzyme reads the opposing strand
sequence and extends the primers by adding nucleotides in the order in which they can pair. • The whole process is repeated over and
over
![Page 9: Molecular marker technology in studies on plant genetic diversity](https://reader036.fdocuments.net/reader036/viewer/2022062412/58838cd41a28ab07628b69db/html5/thumbnails/9.jpg)
• The DNA polymerase, known as 'Taq polymerase', is named after the hot-spring bacterium Thermus aquaticus from which it was originally isolated. • The enzyme can withstand the high temperatures
needed for DNA-strand separation, and can be left in the reaction tube. • The cycle of heating and cooling is repeated over and
over, stimulating the primers to bind to the original sequences and to newly synthesised sequences. • The enzyme will again extend primer sequences. • This cycling of temperatures results in copying and then
copying of copies, and so on, leading to an exponential increase in the number of copies of specific sequences. • Because the amount of DNA placed in the tube at the
beginning is very small, almost all the DNA at the end of the reaction cycles is copied sequences. The reaction products are separated by gel electrophoresis. • Depending on the quantity produced and the size of the
amplified fragment, the reaction products can be visualized directly by staining with ethidium bromide or a silver-staining protocol, or by means of radioisotopes and autoradiography
![Page 10: Molecular marker technology in studies on plant genetic diversity](https://reader036.fdocuments.net/reader036/viewer/2022062412/58838cd41a28ab07628b69db/html5/thumbnails/10.jpg)
PCR procedures: cycle 1
![Page 11: Molecular marker technology in studies on plant genetic diversity](https://reader036.fdocuments.net/reader036/viewer/2022062412/58838cd41a28ab07628b69db/html5/thumbnails/11.jpg)
PCR procedures: cycle 2
![Page 12: Molecular marker technology in studies on plant genetic diversity](https://reader036.fdocuments.net/reader036/viewer/2022062412/58838cd41a28ab07628b69db/html5/thumbnails/12.jpg)
PCR procedures: cycle 3
![Page 13: Molecular marker technology in studies on plant genetic diversity](https://reader036.fdocuments.net/reader036/viewer/2022062412/58838cd41a28ab07628b69db/html5/thumbnails/13.jpg)
CONDITIONSStep Reaction Temperature (°C) Time (Seconds)
1 Initial Denaturation 94 120
2 Cycle Denaturation 94 15
3 Cycle Annealing 58 30
4 Cycle Extension 72 80
5 Repeat from Step 2 to 4 30 Cycles
6 Final extension 72 600
7 Infinite Hold 4
![Page 14: Molecular marker technology in studies on plant genetic diversity](https://reader036.fdocuments.net/reader036/viewer/2022062412/58838cd41a28ab07628b69db/html5/thumbnails/14.jpg)
PCR with arbitrary primers• MAAP (multiple arbitrary amplicon
profiling) is the acronym proposed to cover the three main technologies that fall in this category:• Random amplified polymorphic DNA
(RAPD) • DNA amplification fingerprinting
(DAF) • Arbitrarily primed polymerase chain
reaction (AP-PCR)
![Page 15: Molecular marker technology in studies on plant genetic diversity](https://reader036.fdocuments.net/reader036/viewer/2022062412/58838cd41a28ab07628b69db/html5/thumbnails/15.jpg)
RAPD• Random Amplified Polymorphic DNA• It is a type of PCR reaction, but the segments
of DNA that gets amplified are random. • The researcher performing RAPD creates
several arbitrary, short primers (8–12 nucleotides)
• Then proceeds with the PCR using a large template of genomic DNA, to enable amplification of fragments.
• By resolving the resulting patterns, a semi-unique profile can be gleaned from a RAPD reaction.
• No knowledge of the DNA sequence for the targeted genome is required, as the primers will bind somewhere in the sequence, but it is not certain exactly where.
![Page 16: Molecular marker technology in studies on plant genetic diversity](https://reader036.fdocuments.net/reader036/viewer/2022062412/58838cd41a28ab07628b69db/html5/thumbnails/16.jpg)
• This makes the method popular for comparing the DNA of biological systems that have not had the privileged attention of the scientific community, or in a system in which relatively few DNA sequences are compared (it is not suitable for forming a DNA databank).
• Because it relies on a large, intact DNA template sequence, it has some limitations in the use of degraded DNA samples.
• Its resolving power is much lower than targeted, species specific DNA comparison methods, such as short tandem repeats.
• In recent years, RAPD has been used to characterize, and trace, the phylogeny of diverse plant and animal species.
![Page 17: Molecular marker technology in studies on plant genetic diversity](https://reader036.fdocuments.net/reader036/viewer/2022062412/58838cd41a28ab07628b69db/html5/thumbnails/17.jpg)
How it Works?• Unlike traditional PCR analysis, RAPD does not
require any specific knowledge of the DNA sequence of the target organism: • The identical 1somer primers may or may not
amplify a segment of DNA, depending on positions that are complementary to the primers' sequence. • For example, no fragment is produced if
primers annealed too far apart. • Therefore, if a mutation has occurred in the
template DNA at the site that was previously complementary to the primer, a PCR product will not be produced, resulting in a different pattern of amplified DNA segments on the gel
![Page 18: Molecular marker technology in studies on plant genetic diversity](https://reader036.fdocuments.net/reader036/viewer/2022062412/58838cd41a28ab07628b69db/html5/thumbnails/18.jpg)
Diagrammatic summary
![Page 19: Molecular marker technology in studies on plant genetic diversity](https://reader036.fdocuments.net/reader036/viewer/2022062412/58838cd41a28ab07628b69db/html5/thumbnails/19.jpg)
• This picture shows an image of a very high quality RAPD gel. Both, presence and absence of most bands are very clear and the background is transparent.
• The researcher would have no doubts while selecting bands and collecting data from this gel.
• Consequently, the interpretation of results can be very confident.
![Page 20: Molecular marker technology in studies on plant genetic diversity](https://reader036.fdocuments.net/reader036/viewer/2022062412/58838cd41a28ab07628b69db/html5/thumbnails/20.jpg)
Interpreting RAPD banding patterns
• DNA polymorphism among individuals can be because of:• Mismatches at the primer site• The appearance of a new primer site• The length of the amplified region between primer sites
• Because of the nature of RAPD markers, only the presence or absence of a particular band can be assessed. • Criteria for selecting scoring bands:
• Reproducibility—need to repeat experiments• Thickness• Size• Expected segregation observed in a mapping population
![Page 21: Molecular marker technology in studies on plant genetic diversity](https://reader036.fdocuments.net/reader036/viewer/2022062412/58838cd41a28ab07628b69db/html5/thumbnails/21.jpg)
Examples
Bell pepper
Modern roseMeadow fescue
![Page 22: Molecular marker technology in studies on plant genetic diversity](https://reader036.fdocuments.net/reader036/viewer/2022062412/58838cd41a28ab07628b69db/html5/thumbnails/22.jpg)
Amplified fragment length polymorphisms(AFLPs)
• AFLP is a PCR-based tool used in genetics research, DNA fingerprinting, and in the practice of genetic engineering.
• Developed in the early 1990s by Keygene• AFLP uses restriction enzymes to digest genomic
DNA, followed by ligation of adaptors to the sticky ends of the restriction fragments.
• A subset of the restriction fragments is then selected to be amplified.
• This selection is achieved by using primers complementary to the adaptor sequence, the restriction site sequence and a few nucleotides inside the restriction site fragments.
• The amplified fragments are separated and visualized on polyacrylamide gels, either through autoradiography or fluorescence methodologies, or via automated capillary sequencing instruments
![Page 23: Molecular marker technology in studies on plant genetic diversity](https://reader036.fdocuments.net/reader036/viewer/2022062412/58838cd41a28ab07628b69db/html5/thumbnails/23.jpg)
Digestion
Amplification
Electrophores
isAFLP
The procedure of this technique is divided into three steps:• Digestion of total cellular DNA with one or more
restriction enzymes and ligation of restriction halfsite specific adaptors to all restriction fragments.• Selective amplification of some of these fragments
with two PCR primers that have corresponding adaptor and restriction site specific sequences.• Electrophoretic separation of amplicons on a gel
matrix, followed by visualisation of the band pattern.
![Page 24: Molecular marker technology in studies on plant genetic diversity](https://reader036.fdocuments.net/reader036/viewer/2022062412/58838cd41a28ab07628b69db/html5/thumbnails/24.jpg)
Main features:• A combination of the RFLP and PCR technologies• Based on selective PCR amplification of restriction
fragments from digested DNA• Highly sensitive method for fingerprinting DNA of any
origin and complexity• Can be performed with total genomic DNA or with cDNA
('transcript profiling')• DNA is digested with two different restriction enzymes• Oligonucleotide adapters are ligated to the ends of the
DNA fragments• Specific subsets of DNA digestion products are amplified,
using combinations of selective primers• Polymorphism detection is possible with radioisotopes,
fluorescent dyes or silver staining
![Page 25: Molecular marker technology in studies on plant genetic diversity](https://reader036.fdocuments.net/reader036/viewer/2022062412/58838cd41a28ab07628b69db/html5/thumbnails/25.jpg)
DNA digestion and ligation• One restriction enzyme is a frequent cutter (four-base recognition site, e.g. MseI)
• The second restriction enzyme is a rare cutter (six-base recognition site, e.g. EcoRI)
• Specific synthetic double-stranded adapters for each restriction site are ligated to the DNA fragments generated
![Page 26: Molecular marker technology in studies on plant genetic diversity](https://reader036.fdocuments.net/reader036/viewer/2022062412/58838cd41a28ab07628b69db/html5/thumbnails/26.jpg)
Summarizing the technology
![Page 27: Molecular marker technology in studies on plant genetic diversity](https://reader036.fdocuments.net/reader036/viewer/2022062412/58838cd41a28ab07628b69db/html5/thumbnails/27.jpg)
Interpreting AFLP bands• The AFLP technique detects polymorphisms arising from
changes (presence or size) in the restriction sites or adjacent to these• Different restriction enzymes can be used, and different
combinations of pre- and selective nucleotides willincrease the probability of finding useful polymorphisms• The more selective bases, the less polymorphism will be
detected • Bands are usually scored as either present or absent• Heterozygous versus homozygous bands may be
detected,based on the thickness of the signal, although this can betricky
![Page 28: Molecular marker technology in studies on plant genetic diversity](https://reader036.fdocuments.net/reader036/viewer/2022062412/58838cd41a28ab07628b69db/html5/thumbnails/28.jpg)
Applications• Genetic diversity
assessment• Genetic distance
analysis• Genetic fingerprinting• Analysis of germplasm
collections• Genome mapping• Monitoring diagnostic
markers
Examples
• Limonium sp.• Stylosanthes sp.• Indian mustard
![Page 29: Molecular marker technology in studies on plant genetic diversity](https://reader036.fdocuments.net/reader036/viewer/2022062412/58838cd41a28ab07628b69db/html5/thumbnails/29.jpg)
PCR-based technologies
Sequences-tagged sites
Microsatellites SCARs CAPS ISSRs
![Page 30: Molecular marker technology in studies on plant genetic diversity](https://reader036.fdocuments.net/reader036/viewer/2022062412/58838cd41a28ab07628b69db/html5/thumbnails/30.jpg)
Unlike arbitrary primers, STS rely on some degree of sequence knowledge
Markers based on STS are codominant
They tend to be more reproducible becauselonger primer sequences are used
Require the same basic laboratory protocols and equipment as standard PCR
Sequence-tagged sites as markers
![Page 31: Molecular marker technology in studies on plant genetic diversity](https://reader036.fdocuments.net/reader036/viewer/2022062412/58838cd41a28ab07628b69db/html5/thumbnails/31.jpg)
Microsatellites
• Microsatellites are also called simple sequence repeats (SSRs)• Microsatellites are short tandem repeats (1-
10 bp)• To be used as markers, their location in the
genome of interest must first be identified• Polymorphisms in the repeat region can be
detected by performing a PCR with primers designed from the DNA flanking region
![Page 32: Molecular marker technology in studies on plant genetic diversity](https://reader036.fdocuments.net/reader036/viewer/2022062412/58838cd41a28ab07628b69db/html5/thumbnails/32.jpg)
Identifying microsatellite regions
![Page 33: Molecular marker technology in studies on plant genetic diversity](https://reader036.fdocuments.net/reader036/viewer/2022062412/58838cd41a28ab07628b69db/html5/thumbnails/33.jpg)
Structure
![Page 34: Molecular marker technology in studies on plant genetic diversity](https://reader036.fdocuments.net/reader036/viewer/2022062412/58838cd41a28ab07628b69db/html5/thumbnails/34.jpg)
Selecting primers
![Page 35: Molecular marker technology in studies on plant genetic diversity](https://reader036.fdocuments.net/reader036/viewer/2022062412/58838cd41a28ab07628b69db/html5/thumbnails/35.jpg)
Methodology and visualisation• Methodology:• DNA extraction• PCR with primers specific for microsatellite flanking
regions• Separation of fragments
• Visualization:• By agarose gel electrophoresis, using ethidium
bromide staining and UV light, or • Acrylamide gels using silver staining or radioisotopes,
or • Through automated sequencers, using primers
prelabelled with fluorescence.• Data analysis
![Page 36: Molecular marker technology in studies on plant genetic diversity](https://reader036.fdocuments.net/reader036/viewer/2022062412/58838cd41a28ab07628b69db/html5/thumbnails/36.jpg)
Staining with Ethidium bromide
Staining with Silver Nitrate
![Page 37: Molecular marker technology in studies on plant genetic diversity](https://reader036.fdocuments.net/reader036/viewer/2022062412/58838cd41a28ab07628b69db/html5/thumbnails/37.jpg)
• Advantages:• Require very little and not necessarily high quality
DNA • Highly polymorphic • Evenly distributed throughout the genome • Simple interpretation of results • Easily automated, allowing multiplexing • Good analytical resolution and high
reproducibility
• Disadvantages: • Complex discovery procedure • Costly
![Page 38: Molecular marker technology in studies on plant genetic diversity](https://reader036.fdocuments.net/reader036/viewer/2022062412/58838cd41a28ab07628b69db/html5/thumbnails/38.jpg)
Applications:
Individual genotyping
Germplasm evaluation
Genetic diversity
Genome mapping
Phylogenetic studies
Evolutionary studies
![Page 39: Molecular marker technology in studies on plant genetic diversity](https://reader036.fdocuments.net/reader036/viewer/2022062412/58838cd41a28ab07628b69db/html5/thumbnails/39.jpg)
Sequence characterized amplified
regions (SCARs)
• SCARs take advantage of a band generated through a RAPD experiment
• They use 16-24 bp primers designed from the ends of cloned RAPD markers
• This technique converts a band—prone to difficulties in interpretation and/or reproducibility—into being a very reliable marker
![Page 40: Molecular marker technology in studies on plant genetic diversity](https://reader036.fdocuments.net/reader036/viewer/2022062412/58838cd41a28ab07628b69db/html5/thumbnails/40.jpg)
Steps to obtain SCAR polymorphismsA potentially
interesting band is
identified in a RAPD gel
The band is cut out of the gelThe DNA fragment
is cloned in a vector and sequenced
Specific primers (16-24 bp long) for that DNA fragment are designed
Re-amplification of the template DNA
with the new primers will show a new and simpler PCR pattern
![Page 41: Molecular marker technology in studies on plant genetic diversity](https://reader036.fdocuments.net/reader036/viewer/2022062412/58838cd41a28ab07628b69db/html5/thumbnails/41.jpg)
Diagram of the SCAR procedure
![Page 42: Molecular marker technology in studies on plant genetic diversity](https://reader036.fdocuments.net/reader036/viewer/2022062412/58838cd41a28ab07628b69db/html5/thumbnails/42.jpg)
Advantages Disadvantages
Simpler patterns than RAPDs
Robust assay due to
the design of specific long
primers
Mendelian inheritance.
Sometimes convertible
to codominant markers
Require at least a small
degree of sequence knowledge
Require effort and
expense in designing
specific primers for each locus
![Page 43: Molecular marker technology in studies on plant genetic diversity](https://reader036.fdocuments.net/reader036/viewer/2022062412/58838cd41a28ab07628b69db/html5/thumbnails/43.jpg)
Cleaved amplified polymorphicsequence (CAPS)
• This method is based on • The design of specific primers, • Amplification of DNA fragments, and • Generation of smaller, possibly variable,
fragments by means of a restriction enzyme• This technique aims to convert an
amplified band that does not show variation into a polymorphic one
![Page 44: Molecular marker technology in studies on plant genetic diversity](https://reader036.fdocuments.net/reader036/viewer/2022062412/58838cd41a28ab07628b69db/html5/thumbnails/44.jpg)
Steps for generating CAPSA band, DNA, gene sequence
or other type of marker isidentified as important
Either the band is detected through PCR (and cut out of
the gel, and the fragment cloned and sequenced) or the fragment sequence is already
available
Specific primers are designed from the fragment
sequence
The newly designed primers are used to amplify the
template DNA
The PCR product is subjected to digestion by a
panel ofrestriction enzymes
Polymorphism may be identified with some of the
enzymes
![Page 45: Molecular marker technology in studies on plant genetic diversity](https://reader036.fdocuments.net/reader036/viewer/2022062412/58838cd41a28ab07628b69db/html5/thumbnails/45.jpg)
Generating CAPS
![Page 46: Molecular marker technology in studies on plant genetic diversity](https://reader036.fdocuments.net/reader036/viewer/2022062412/58838cd41a28ab07628b69db/html5/thumbnails/46.jpg)
Advantages
• Robust assay
because specific long
primers are designed
• Codominant markers
• Benefit from markers
that may have
already been
mapped
• Identify
polymorphisms in
markers that were
previously not
informative
Disadvantages
• Require at least a
small amount of
sequence knowledge
• Effort and expense
required to design
specific primers
for each locus
![Page 47: Molecular marker technology in studies on plant genetic diversity](https://reader036.fdocuments.net/reader036/viewer/2022062412/58838cd41a28ab07628b69db/html5/thumbnails/47.jpg)
Inter-simple sequence repeats (ISSRs)
• They are regions found between microsatellite repeats• The technique is based on PCR
amplification ofintermicrosatellite sequences• Because of the known abundance of
repeat sequences spread all over the genome, it targets multiple loci
![Page 48: Molecular marker technology in studies on plant genetic diversity](https://reader036.fdocuments.net/reader036/viewer/2022062412/58838cd41a28ab07628b69db/html5/thumbnails/48.jpg)
Identifying ISSR polymorphisms
• A typical PCR is performed in which primers have been designed, based on a microsatellite repeat sequence, and extended one to several bases into the flanking sequence as anchor points. • Different alternatives are possible:
• Only one primer is used• Two primers of similar characteristics are used• Combinations of a microsatellite-sequence
anchored primer with a random primer (i.e. those used for RAPD)
![Page 49: Molecular marker technology in studies on plant genetic diversity](https://reader036.fdocuments.net/reader036/viewer/2022062412/58838cd41a28ab07628b69db/html5/thumbnails/49.jpg)
Designing primers for ISSR polymorphisms
![Page 50: Molecular marker technology in studies on plant genetic diversity](https://reader036.fdocuments.net/reader036/viewer/2022062412/58838cd41a28ab07628b69db/html5/thumbnails/50.jpg)
• The diagram above presents three different items:
• The original DNA sequence in which two different repeated sequences (CA), inversely oriented, are identified. Both repeated sections are, in addition, closely spaced.
• If primers were designed from within the repeated region only, the interrepeat section would be amplified but locus-specificity might not be guaranteed. In the second row, a PCR product is shown as a result of amplification from a 3'-anchored primer (CA)n NN at each end of the interrepeat region. CA is the repeat sequence that was extended by NN, two nucleotides running into the interrepeat region.
• Alternatively, anchors may be chosen from the 5' region. The PCR product in the third row is a result of using primers based on the CA repeat but extended at the 5' end by NNN and NN, respectively
![Page 51: Molecular marker technology in studies on plant genetic diversity](https://reader036.fdocuments.net/reader036/viewer/2022062412/58838cd41a28ab07628b69db/html5/thumbnails/51.jpg)
Advantages• Do not require prior sequence
information• Variation within unique regions of
the genome may be found at several loci simultaneously
• Tend to identify significant levels of variation
• Microsatellite sequence-specific• Very useful for DNA profiling,
especially for closely related species
Disadvantages• Dominant markers• Polyacrylamide gel
electrophoresis and detection with silver staining or radioisotopes may be needed
![Page 52: Molecular marker technology in studies on plant genetic diversity](https://reader036.fdocuments.net/reader036/viewer/2022062412/58838cd41a28ab07628b69db/html5/thumbnails/52.jpg)
Latest strategies
DNA sequencing Expressed sequence tags (ESTs)
Microarray technology
Diversity array technology (DArT)
Single nucleotide polymorphisms
(SNPs)
![Page 53: Molecular marker technology in studies on plant genetic diversity](https://reader036.fdocuments.net/reader036/viewer/2022062412/58838cd41a28ab07628b69db/html5/thumbnails/53.jpg)
References…
References
Plant Biotechnology and Molecular
Markers• P.S. Srivastava• Alka Narula
• Sheela Srivastava
Using molecular marker
technology in studies on plant genetic diversity• M. Carmen de
Vicente• Theresa Fulton
Molecular Markers
• Ashwani Kumar• Scientific Blogging 2.0
![Page 54: Molecular marker technology in studies on plant genetic diversity](https://reader036.fdocuments.net/reader036/viewer/2022062412/58838cd41a28ab07628b69db/html5/thumbnails/54.jpg)