Simple and robust screening of pooled yeast artificial chromosome libraries by the restriction enzyme digestion of polymerase chain reaction products

Xueqian Wang, Jing Qian, Minoru S.H. Ko

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Abstract

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.

Original languageEnglish
Pages (from-to)63-68
Number of pages6
JournalGenetic Analysis: Biomolecular Engineering
Volume11
Issue number3
DOIs
Publication statusPublished - 1994
Externally publishedYes

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ASJC Scopus subject areas

  • Applied Microbiology and Biotechnology
  • Genetics

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