cDNA selection with YACs.

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.

journal_name

Mol Biotechnol

journal_title

Molecular biotechnology

authors

Parimoo S

doi

10.1007/BF02760779

subject

Has Abstract

pub_date

1997-12-01 00:00:00

pages

255-68

issue

3

eissn

1073-6085

issn

1559-0305

journal_volume

8

pub_type

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