Predigestion of DNA template improves the level of polymorphism of random amplified polymorphic DNAs in wheat.
Random amplified polymorphic DNA (RAPD) analysis in wheat has proven to be poor in its levels of both reproducibility and polymorphism. By digesting the template, prior to performing PCR, with frequently cutting restriction enzymes, the level of polymorphism was improved. RAPD profiles from certain primers were not affected by this pretreatment of the template, but other primers produced distinct profiles from each of several restriction enzymes assayed. Some polymorphisms were specific to one or more restriction digests, but none involved the simple loss of bands from the unrestricted template profile. Genotypic comparisons enabled the selection of primer-restriction enzyme combinations that enabled polymorphic and mappable patterns to be produced both between wheat varieties and between wheats with and without chromosomal segments deriving from related species.
Koebner
R M
RM
John Innes Centre, Colney, Norwich, UK.
D017931:DNA Primers; D004262:DNA Restriction Enzymes; D018744:DNA, Plant / Q000378:metabolism*; D005838:Genotype; D018515:Plant Leaves; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D013698:Templates, Genetic; D014908:Triticum / Q000235:genetics*
D017931:DNA Primers; D018744:DNA, Plant; D004262:DNA Restriction Enzymes