Cloning multigene families with degenerate PCR primers

Thomas M. Wilkie; Melvin I. Simon

doi:10.1016/S1046-2023(05)80123-9

Cloning multigene families with degenerate PCR primers

Thomas M. Wilkie, Melvin I. Simon

Research output: Contribution to journal › Article › peer-review

32 Scopus citations

Abstract

Multigene families often exhibit regions of highly conserved amino acid sequence that are characteristic of a given family. Degenerate PCR primers that target these motifs will selectively amplify members within that multigene family. Members of the family that are expressed in a particular tissue can be amplified from cDNA. The PCR products can be easily cloned, screened, and sequenced over a precise region to rapidly identify novel genes. PCR with degenerate primers can also be used to screen the expression of these genes in various tissues and cell types.

Original language	English (US)
Pages (from-to)	32-41
Number of pages	10
Journal	Methods
Volume	2
Issue number	1
DOIs	https://doi.org/10.1016/S1046-2023(05)80123-9
State	Published - Feb 1991

ASJC Scopus subject areas

Molecular Biology
General Biochemistry, Genetics and Molecular Biology

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10.1016/S1046-2023(05)80123-9

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title = "Cloning multigene families with degenerate PCR primers",

abstract = "Multigene families often exhibit regions of highly conserved amino acid sequence that are characteristic of a given family. Degenerate PCR primers that target these motifs will selectively amplify members within that multigene family. Members of the family that are expressed in a particular tissue can be amplified from cDNA. The PCR products can be easily cloned, screened, and sequenced over a precise region to rapidly identify novel genes. PCR with degenerate primers can also be used to screen the expression of these genes in various tissues and cell types.",

author = "Wilkie, {Thomas M.} and Simon, {Melvin I.}",

note = "Funding Information: We thank Mike Strathmann for discussions and numerous contributions, Carol Lee for screening and sequencing the PCR clones, Anthony Bellv~ for isolating the spermatogenic cell types, members of the Microchemical Facility at Caltech for oligonucleotide synthesis and for coupling inosine to columns on special order, Pantelis Tsoulfas for TFASTA sequence comparisons, and Jane Johnson, Murray Robinson, and Pantelis Tsoulfas for comments on the manuscript. This work was supported in part by a NIH postdoctoral fellowship to T.M.W. (GMl1576) and a NIH grant to M.I.S. (GM34236).",

year = "1991",

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doi = "10.1016/S1046-2023(05)80123-9",

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AU - Simon, Melvin I.

N1 - Funding Information: We thank Mike Strathmann for discussions and numerous contributions, Carol Lee for screening and sequencing the PCR clones, Anthony Bellv~ for isolating the spermatogenic cell types, members of the Microchemical Facility at Caltech for oligonucleotide synthesis and for coupling inosine to columns on special order, Pantelis Tsoulfas for TFASTA sequence comparisons, and Jane Johnson, Murray Robinson, and Pantelis Tsoulfas for comments on the manuscript. This work was supported in part by a NIH postdoctoral fellowship to T.M.W. (GMl1576) and a NIH grant to M.I.S. (GM34236).

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Y1 - 1991/2

N2 - Multigene families often exhibit regions of highly conserved amino acid sequence that are characteristic of a given family. Degenerate PCR primers that target these motifs will selectively amplify members within that multigene family. Members of the family that are expressed in a particular tissue can be amplified from cDNA. The PCR products can be easily cloned, screened, and sequenced over a precise region to rapidly identify novel genes. PCR with degenerate primers can also be used to screen the expression of these genes in various tissues and cell types.

AB - Multigene families often exhibit regions of highly conserved amino acid sequence that are characteristic of a given family. Degenerate PCR primers that target these motifs will selectively amplify members within that multigene family. Members of the family that are expressed in a particular tissue can be amplified from cDNA. The PCR products can be easily cloned, screened, and sequenced over a precise region to rapidly identify novel genes. PCR with degenerate primers can also be used to screen the expression of these genes in various tissues and cell types.

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Cloning multigene families with degenerate PCR primers

Abstract

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