Specific DNA amplification utilizing the polymerase chain reaction and random oligonucleotide primers: application to the analysis of antigen receptor variable regions

DNA Cell Biol. 1992 Nov;11(9):707-20. doi: 10.1089/dna.1992.11.707.

Abstract

The polymerase chain reaction (PCR) allows rapid amplification of DNA of known sequence. In many situations, part of a genetic sequence is known, but adjacent sequences of interest are unknown. This is common in investigations of antigen receptor genes from B and T lymphocytes, which are composed of a constant region of known sequence and a variable region, for which the sequence may not be known. Herein is described a method to amplify DNA when sequence information is available for only one primer. This procedure utilizes a primer of known sequence in conjunction with a mixture of short random primers. Application of this method to the amplification of T-cell antigen receptor cDNA is described.

Publication types

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

MeSH terms

  • Arthritis, Rheumatoid / immunology
  • Base Sequence
  • Cells, Cultured
  • Cloning, Molecular
  • DNA, Single-Stranded
  • Humans
  • Molecular Sequence Data
  • Oligodeoxyribonucleotides
  • Polymerase Chain Reaction / methods*
  • Receptors, Antigen, T-Cell / genetics*
  • Synovial Fluid / cytology
  • T-Lymphocytes / metabolism
  • Templates, Genetic

Substances

  • DNA, Single-Stranded
  • Oligodeoxyribonucleotides
  • Receptors, Antigen, T-Cell