Analysis of DNA from A Single Barley Embryo by the DNA Polymerase Chain Reaction (PCR) Technique

Abstract

This study was conducted to investigate whether DNA extracted from a single barley embryo is suitable for conducting polymerase chain reaction (PCR) amplification. We extracted DNA from one-seed and five-seed samples of four barley cultivars, ‘Bowman’, ‘Colter’, ‘Crystal’, and ‘Russell’. Six random-amplified polymorphic DNA (RAPD) primers and one sequence-tagged-site (STS) primer pair were tested for DNA amplification using both Taq DNA polymerase and the Stoffel fragment, which is a modified form of recombinant Taq DNA polymerase. DNA polymorphism was found with the STS primer and five of the RAPD primers. The banding patterns of DNA from one-seed samples were almost identical to those of the five-seed samples. Additional testing of nine barley cultivars was conducted to compare embryo and leaf tissue DNA PCR amplification results. Our tests indicated that DNA extracted from a single embryo is practical for PCR analysis. The technique we utilized is simple, fast, and can be applied to the identification of barley cultivars.