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Exons – Introns = Lexons: In-frame concatenation of exons by PCR

✍ Scribed by Thérèse M. F. Tuohy; Joanna Groden

Book ID: 101259486
Publisher: John Wiley and Sons
Year: 1998
Tongue: English
Weight: 324 KB
Volume: 12
Category: Article
ISSN: 1059-7794
DOI: 10.1002/(sici)1098-1004(1998)12:2<122::aid-humu7>3.0.co;2-w

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✦ Synopsis

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 intronexon 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) spliceforms to be amplified. We have used this technique to obtain concatamers of exons 39A 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.

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