Single-step protocol for preparation of plant tissue for analysis by PCR.
PCR has many applications in the isolation and analysis of plant DNA. The influence of salt and EDTA concentration, pH, incubation time and temperature on the preparation of plant material for PCR was evaluated. A general single-step method was developed in which a small amount of plant tissue was heated in a simple solution. The DNA in the supernatant was found to be suitable for most PCR applications including arbitrarily primed PCR (random-amplified polymorphic DNA) and PCR with specific primers for both single- and multiple-copy genes. The technique is much simpler than those generally used for plant DNA preparation and was successful with tissues from a wide range of species.
D001483:Base Sequence; D018744:DNA, Plant / Q000032:analysis*; D004492:Edetic Acid / Q000494:pharmacology; D002523:Edible Grain / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D018515:Plant Leaves / Q000737:chemistry; D010944:Plants / Q000235:genetics*; D016133:Polymerase Chain Reaction; D011189:Potassium Chloride / Q000494:pharmacology; D012639:Seeds / Q000737:chemistry; D013696:Temperature; D013997:Time Factors
D018744:DNA, Plant; D011189:Potassium Chloride; D004492:Edetic Acid