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

Biotechniques. 1996 Oct;21(4):644-6, 648-9. doi: 10.2144/96214st01.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • DNA, Complementary / chemistry*
  • Humans
  • Molecular Sequence Data
  • Polymerase Chain Reaction / methods*
  • RNA, Messenger / analysis*

Substances

  • DNA, Complementary
  • RNA, Messenger

Associated data

  • GENBANK/UNKNOWN

Grants and funding