Abstract
Inter-simple sequence repeat PCR (ISSR-PCR) is a fast, inexpensive genotyping technique based on length variation in the regions between microsatellites. The method requires no species-specific prior knowledge of microsatellite location or composition. Very small amounts of DNA are required, making this method ideal for organisms of conservation concern, or where the quantity of DNA is extremely limited due to organism size. ISSR-PCR can be highly reproducible but requires careful attention to detail. Optimization of DNA extraction, fragment amplification, and normalization of fragment peak heights during fluorescent detection are critical steps to minimizing the downstream time spent verifying and scoring the data.
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Acknowledgements
I am grateful to the team at Applied Biosystems, Inc. (C.J. Davidson and T. Ingalls) for collaborative efforts to optimize ISSR methods on the 3500xL Genetic Analyzer, and to Rancho Santa Ana Botanic Garden for financial support.
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Prince, L.M. (2015). Plant Genotyping Using Fluorescently Tagged Inter-Simple Sequence Repeats (ISSRs): Basic Principles and Methodology. In: Batley, J. (eds) Plant Genotyping. Methods in Molecular Biology, vol 1245. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1966-6_5
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DOI: https://doi.org/10.1007/978-1-4939-1966-6_5
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