Plant genotyping using fluorescently tagged inter-simple sequence repeats (ISSRs): basic principles and methodology.
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.
Department of Botany, The Field Museum, 1400 S Lake Shore Drive, Chicago, IL, 60605, USA, lprince@fieldmuseum.org.
Linda M
LM
Prince
D020029:Base Pairing / Q000235:genetics; D018744:DNA, Plant / Q000302:isolation & purification; D005456:Fluorescent Dyes / Q000378:metabolism*; D060005:Genotyping Techniques / Q000379:methods*; D018895:Microsatellite Repeats / Q000235:genetics*; D010944:Plants / Q000235:genetics*; D016133:Polymerase Chain Reaction
D018744:DNA, Plant; D005456:Fluorescent Dyes