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Inter Sequence Specific Repeat (ISSR) PCR (CAT#: STEM-MB-0207-WXH)

Introduction

Inter simple sequence repeat (ISSR)-PCR is a technique that involves the use of microsatellite sequences as primers in a polymerase chain reaction to generate multi-locus markers.<br />The main advantage of ISSRs is that no sequence data for primer construction are needed. Because the analytical procedures include PCR, only low quantities of template DNA are required.<br />It is a simple and quick method that combines most of the advantages of microsatellites (SSRs) and amplified fragment length polymorphism (AFLP) with the universality of RAPD.




Principle

ISSRs are DNA fragments of about 100-3000 bp located between adjacent, oppositely oriented microsatellite regions. ISSRs are amplified by PCR using microsatellite core sequences as primers with a few selective nucleotides as anchors into the non-repeat adjacent regions (16-18 bp). About 10-60 fragments from multiple loci are generated simultaneously, separated by gel electrophoresis and scored as the presence or absence of fragments of particular size.

Applications

• Genomic fingerprinting
• Genetic diversity and phylogenetic analysis
• Genome mapping
• Gene tagging

Procedure

1.Primer design
2.DNA Extraction and purification
3.ISSR-PCR
4.Data analysis

Materials

ISSR markers