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Semi-multiplex PCR technique for screening of abundant transcripts during systematic sequencing of cDNA libraries
B Pacchioni1, S Trevisan, S Gomirato
1CRIBI Biotechnology Centre, Università degli Studi di Padova, Italy.
Abstract:
The systematic sequencing of cDNA libraries is an efficient approach for the identification of new genes, but the presence of abundant mRNAs is often a major problem. This paper describes a very simple method of "semi-multiplex PCR" that allows specific identification of such abundant transcripts before DNA sequencing without using nonrepresentative subtracted libraries. The PCR utilizes a series of forward primers specific for abundant transcripts with a pair of universal primers used for template generation. cDNA clones corresponding to abundant mRNAs are then revealed by double bands in agarose gel.
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