Simple and robust screening of pooled yeast artificial chromosome libraries by the restriction enzyme digestion of polymerase chain reaction products.
The polymerase chain reaction (PCR)-based method for screening pooled yeast artificial chromosome (YAC) libraries was modified by adding a step for the restriction enzyme digestion of the PCR products. This modification significantly increased the reliability of YAC screening and made it possible to identify YAC clones without a cumbersome verification step by colony hybridization and/or Southern blotting. Using this method, we assigned 39 mouse YAC clones to a mouse genetic map with 13 biallelic, polymorphic expressed sequence tags. This method provides a fast, reliable way to identify YAC clones with PCR-based sequence-tagged sites.
Wang
X
X
Center for Molecular Medicine and Genetics, Wayne State University School of Medicine, Detroit, Michigan.
Qian
J
J
Ko
M S
MS
D000818:Animals; D001483:Base Sequence; D018244:Chromosomes, Artificial, Yeast; D017931:DNA Primers; D018076:DNA, Complementary; D015870:Gene Expression; D015698:Genomic Library; D051379:Mice; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D015183:Restriction Mapping
D017931:DNA Primers; D018076:DNA, Complementary