METHOD FOR AMPLIFICATION OF DNA FRAGMENT

摘要

Disclosed is a method for the amplification of a DNA fragment, which is intended to be used in a transposon display method for a higher plant, which can amplify only a labeled PCR product selectively, and which shows little variations in experimental results among individuals. The method for the amplification of a DNA fragment comprises the following steps in this order: a DNA extraction step for extracting genomic DNA from a cell of a higher plant; an adapter addition step for partially digesting the genomic DNA with a restriction enzyme to produce a DNA fragment and then adding an adapter to the DNA fragment; a first PCR step for amplifying the DNA fragment by an one-sided PCR procedure which uses a DNA fragment ligated to the adapter as a template and also uses only a primer specific to the transposon; and a second PCR step for amplifying the DNA fragment by a two-sided PCR procedure which uses a DNA fragment amplified in the first PCR step as a template and also uses both a labeled primer specific to the transposon and a primer complementary to the adapter.