Semi-multiplex PCR technique for screening of abundant transcripts during systematic sequencing of cDNA libraries.

نویسندگان

  • B Pacchioni
  • S Trevisan
  • S Gomirato
  • S Toppo
  • G Valle
  • G Lanfranchi
چکیده

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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عنوان ژورنال:
  • BioTechniques

دوره 21 4  شماره 

صفحات  -

تاریخ انتشار 1996