A lock-docking oligo(dT) primer for 5' and 3' RACE PCR

PCR Methods Appl. 1992 Nov;2(2):144-8. doi: 10.1101/gr.2.2.144.

Abstract

We describe a method that can be used to obtain and sequence 3' and 5' ends of cDNA transcripts directly from PCR products. The method employs a modified oligo(dT) primer that enables it to "lock-dock" at the junction of gene-specific cDNA sequence and a natural (3') or appended (5') poly(A) tail. As a result, discrete, first-round PCR products are obtained that are easily isolated and sequenced directly.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Base Sequence
  • DNA / genetics
  • Dogs
  • Glucose Transporter Type 3
  • Molecular Sequence Data
  • Monosaccharide Transport Proteins / genetics
  • Nerve Tissue Proteins*
  • Poly T*
  • Polymerase Chain Reaction / methods*

Substances

  • Glucose Transporter Type 3
  • Monosaccharide Transport Proteins
  • Nerve Tissue Proteins
  • Poly T
  • DNA