Open AccessApplication of In Situ PCR to Yeast Cells for Screening YAC Libraries
Author(s) -
Saika Aytay,
Lisa M. Davis,
John Hozier
Publication year - 1999
Publication title -
biotechniques/biotechniques
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.617
H-Index - 131
eISSN - 1940-9818
pISSN - 0736-6205
DOI - 10.2144/99273st06
Subject(s) - yeast artificial chromosome , genomic library , yeast , biology , microbiology and biotechnology , cloning (programming) , genetics , computational biology , polymerase chain reaction , chromosome , in situ , molecular cloning , genomic dna , microsatellite , gene mapping , dna , gene , chemistry , base sequence , computer science , peptide sequence , allele , programming language , organic chemistry
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 solutionbased PCR screening. This approach can conceivably be applied to the products of automated robotic workstations.