cDNA selection with YACs

Molecular Biotechnology
S Parimoo

Abstract

Identification of expressed sequence tags (ESTs) in large genomic segments is an important step in positional cloning and genomic mapping studies. A simple and efficient polymerase chain reaction (PCR)-based approach is described here to identify coding sequences in large genomic fragments of DNA cloned in vectors such as yeast artificial chromosome (YAC) vectors. The method is based on blocking of sequences such as repetitive and GC rich sequences in the genomic DNA immobilized on nylon paper discs prior to hybridization of the discs to cDNA library, and recovery of the selected cDNAs by the PCR. Single or multiple cDNA libraries can be used in the selection procedure. The procedure has been used successfully also with total yeast DNA containing a YAC.

References

May 1, 1991·Proceedings of the National Academy of Sciences of the United States of America·A J BucklerD E Housman
Nov 1, 1991·Proceedings of the National Academy of Sciences of the United States of America·S ParimooS M Weissman
Jun 21, 1991·Science·H A ErlichJ J Sninsky
Jul 11, 1990·Nucleic Acids Research·P ElvinA F Markham
Nov 1, 1990·Proceedings of the National Academy of Sciences of the United States of America·G M DuykD R Cox
Mar 1, 1985·Proceedings of the National Academy of Sciences of the United States of America·W I WoodR M Lawn
Dec 15, 1993·Proceedings of the National Academy of Sciences of the United States of America·H WeiS M Weissman
Sep 11, 1993·Nucleic Acids Research·S ParimooS M Weissman

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