Application of in situ PCR to yeast cells for screening YAC libraries

We have used in situ PCR technology in yeast cells with the ultimate goal o f cloning and screening genomic yeast artificial chromosome (YAC) libraries The target sequences in YAC clones were amplified "in situ" in yeast cells by the same set of microsatellite primers used in solution-based PCR scree ning. The method is fast and sensitive and obviates the steps required for individual isolation of DNAs from hundreds to thousands of YAC clones and t hus has an advantage over conventional solution based PCR screening. This a pproach can conceivably be applied to the products of automated robotic wor kstations.