Ultra-simple DNA extraction method for marker-assisted selection using microsatellite markers in rice

We present an ultra-simple DNA extraction method for microsatellite analysi s of rice. Each extraction requires only one microtube, one disposable pipe tte tip, TE buffer and few pieces (about 5 mm) of rice leaf tissue. This is sufficient for 200 PCR reactions. The extract can be kept in the freezer f or long-term storage. Also, DNA can be extracted from 200-300 individuals i n a few hours. These features enabled us to perform rapid large-scale seedl ing genotyping required for marker-assisted selection. We have also examine d the applicability of this method for other PCR-based markers: RAPDs, nucl ear STS, chloroplast STS and chloroplast microsatellites.