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 of 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 screening. The method is fast and sensitive and obviates the steps required for individual isolation of DNAs from hundreds to thousands of YAC clones and thus has an advantage over conventional solution-based PCR screening. This approach can conceivably be applied to the products of automated robotic workstations.
Aytay
S
S
Applied Genetics Laboratories, Melbourne, FL, USA.
Davis
L M
LM
Hozier
J C
JC
D000818:Animals; D001322:Autoanalysis; D018244:Chromosomes, Artificial, Yeast; D017931:DNA Primers; D004271:DNA, Fungal / Q000302:isolation & purification*; D015723:Gene Library; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D051379:Mice; D008810:Mice, Inbred C57BL; D018895:Microsatellite Repeats
D017931:DNA Primers; D004271:DNA, Fungal