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Plant genotyping using fluorescently tagged inter-simple sequence repeats (ISSRs): basic principles and methodology
1Department of Botany, The Field Museum, 1400 S Lake Shore Drive, Chicago, IL, 60605, USA, lprince@fieldmuseum.org.
Methods in Molecular Biology (Clifton, N.J.)
|November 7, 2014
Summary
Inter-simple sequence repeat PCR (ISSR-PCR) offers a rapid, cost-effective genotyping method ideal for limited DNA samples. Careful optimization ensures high reproducibility for genetic studies, especially in conservation biology.
Area of Science:
- Genetics
- Molecular Biology
- Conservation Biology
Background:
- Microsatellites are repetitive DNA sequences.
- Length variation in inter-microsatellite regions can be used for genotyping.
Purpose of the Study:
- To describe Inter-simple sequence repeat PCR (ISSR-PCR) as a genotyping technique.
- To highlight its advantages for specific applications.
Main Methods:
- Utilizes length polymorphism in DNA regions between microsatellites.
- Requires minimal species-specific prior knowledge.
- Demands small DNA quantities.
Main Results:
- ISSR-PCR is fast and inexpensive.
- It is suitable for organisms with limited DNA availability, such as those of conservation concern.
- The method can be highly reproducible with careful optimization.
Conclusions:
- ISSR-PCR is a valuable genotyping tool, particularly for conservation genetics.
- Optimization of DNA extraction, amplification, and data normalization is crucial for reliable results.
