TY - JOUR
T1 - Simple and robust screening of pooled yeast artificial chromosome libraries by the restriction enzyme digestion of polymerase chain reaction products
AU - Wang, Xueqian
AU - Qian, Jing
AU - Ko, Minoru S.H.
N1 - Funding Information:
We thank Shirley Tilghman and David Koos for providing pooled mouse YAC libraries and for discussion, and Yoshio Sugimura for customizing a polyacrylamide gel electrophoresis system. We also thank Craig N. Giroux and Suman L. Misra for the instruction of yeast cell culture, and Orlando J. Miller for the critical reading of the manuscript. This research was partially supported by the State of Michigan Research Excellence Fund through the Center for Molecular Biology, Wayne State University, and by Daiichi Pharmaceutical Co. Ltd. J.Q. was supported by NIH grant (RR03332) to J. Christopher Stotes.
PY - 1994
Y1 - 1994
N2 - 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 sequenc-tagged sites.
AB - 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 sequenc-tagged sites.
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U2 - 10.1016/1050-3862(94)90052-3
DO - 10.1016/1050-3862(94)90052-3
M3 - Article
C2 - 7803131
AN - SCOPUS:0028028549
SN - 1871-6784
VL - 11
SP - 63
EP - 68
JO - Biomolecular Engineering
JF - Biomolecular Engineering
IS - 3
ER -