A rapid and reliable DNA preparation method for screening a large number of yeast clones by polymerase chain reaction

摘要

Yeast is an ideal host system for studying exogenous eukaryotic gene expression and for studying gene structures from recombinant plasmid DNAs and yeast artificial chromosomes (YACs). Many yeast expression studies may also require the altering of thegenome itself by homologous recombination. All these studies require the screening for positive transformants and the most popularly used method of screening is PCR. As some gene replacement experiments have a low success rate (as low as 4%) (1), it becomes necessary to screen a large number of clones. Hundreds and thousands of transformants, e.g. 1200 YAC clones/ day/lab as reported by Khristich et al. (2), are screened by PCR in many laboratories.