酿酒酵母染色体DNA样品制备和CHEF分析

PREPARATION AND CHEF ANALYSIS OF YEAST CHROMOSOMAL DNA SAMPLE

等高锁状均匀电场（contolllsclampedhomogeneouselectncfield，CHED电泳技术与分子杂交、酵母人工染色体（yeastartificialchromosome，YAC）等分子生物学技术的有机结合，推动了真菌电泳核型、染色体作图和染色体DNA长度多型性（chromosomalDNAlellgthpolymorphism，CLP...

1.5% Lywallzyme (product of Guangdong Microbiology Institute, China) was used to remove the cell walls of Saccharomyces cerevisae in the preparation of protoplasts. The protoplasts resuspented in ST buffer at the concentration of 4 × 108 / ml were mixed with an equal volume of 1.0% low-melting point agarose in ST buffer at 50℃. In order to prepare yeast chromosomal DNA samples, the chilled agarose Plugs were incubated in NDS buffer containing ling/ml proteinase K for 24h. This procedure of chromosomal DNA sample prepeon,which is different from that adopted by former researchers, have laid a fundation for the preparation of megabase-size DNA marker and electrophorehc karyotype analysis of other fungi. It was revealed by CHEF electrophoresis that chromosomal DNA length polymorphism exists between tested stain X8 and control stain ThH755.