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PMID: 1946377 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

cDNA selection: efficient PCR approach for the selection of cDNAs encoded in large chromosomal DNA fragments.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 88 ·No. 21 ·1991-11-01 ·Pages 9623-7

Parimoo S, Patanjali SR, Shukla H, Chaplin DD, Weissman SM

Abstract

Identification of coding segments in large fragments of genomic DNA is a recurrent problem in genome mapping and positional cloning studies. We have developed a rapid and efficient protocol to achieve this goal, based on hybridization of cDNA fragments to immobilized DNA and recovery of the selected cDNAs by the PCR. The procedure permits rapid cloning of cDNA fragments encoded by large genomic DNA fragments, groups of yeast artificial chromosomes, or cosmids and has the potential to directly enrich cDNAs encoded in chromosome segments. By this approach we have been able to identify several non-major histocompatibility complex class I clones from a yeast artificial chromosome that includes the HLA-A locus.

MeSH Terms
Base Sequence Cloning, Molecular/methods DNA/genetics Gene Library Genetic Vectors Genomic Library Humans Molecular Sequence Data Nucleic Acid Hybridization Oligonucleotides/chemistry Polymerase Chain Reaction/methods Repetitive Sequences, Nucleic Acid
Chemicals
Oligonucleotides DNA
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Parimoo S
Department of Genetics, Yale University School of Medicine, New Haven, CT 06510.
Patanjali S R
Shukla H
Chaplin D D
Weissman S M
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.88.21.9623
Published
1991-11-01
Pages
9623-7
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC52770
Subset
IM
Grants
NIAID NIH HHS · AI15322 · United States
NHGRI NIH HHS · P50 HG00201 · United States
NCI NIH HHS · R35 CA42556-06 · United States
External Links
PubMed source DOI: 10.1073/pnas.88.21.9623 PMC Full Text View journal details
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