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Exons – Introns = Lexons: In‐frame concatenation of exons by PCR
Author(s) -
Tuohy Thérèse M. F.,
Groden Joanna
Publication year - 1998
Publication title -
human mutation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.981
H-Index - 162
eISSN - 1098-1004
pISSN - 1059-7794
DOI - 10.1002/(sici)1098-1004(1998)12:2<122::aid-humu7>3.0.co;2-w
Subject(s) - exon , biology , intron , genetics , genomic dna , concatenation (mathematics) , exon trapping , complementary dna , tandem exon duplication , splice , gene , microbiology and biotechnology , computational biology , alternative splicing , mathematics , combinatorics
A method for concatenating exons from genomic DNA, thereby skipping large stretches of intron sequence, has been developed using the polymerase chain reaction (PCR) with primers based on known intron–exon junction sequences. The use of genomic DNA circumvents the need for cDNA preparation for many purposes, including cDNA construction and mutational analysis. This PCR method also facilitates the concatenation of nonconsecutive exons, allowing different (known or hypothetical) splice‐forms to be amplified. We have used this technique to obtain concatamers of exons 3–9A of APC , a tumor suppressor gene that is mutated in sporadic colorectal cancers and in the germline of individuals with adenomatous polyposis coli. This method also facilitates the generation of any polymorphic derivative of a known sequence, even where the derivative differs from the available sequence at several positions. Hum Mutat 12:122–127, 1998. © 1998 Wiley‐Liss, Inc.