Molecular Biotechnology

, Volume 3, Issue 2, pp 129–134

Selection of primers for polymerase chain reaction

  • Wojciech Rychlik
Protocol

DOI: 10.1007/BF02789108

Cite this article as:
Rychlik, W. Mol Biotechnol (1995) 3: 129. doi:10.1007/BF02789108

Abstract

One of the most important factors affecting the quality of PCR is the choice of primers. In general, the longer the PCR product the more difficult it is to select efficient primers and set appropriate designing primers, and in general, the more DNA sequence information is available, the better the ch0ance of finding an optimal primer pair. Efficient primers can be designed by avoiding the following flaws: primer-dimer formation, self-complementarity, too lowTm of the primers, and/or their incorrect internal stability profile. Tips on subcloning PCR products, calculating duplex stability (predicting dimer formation strength), and designing degenerate primers are given.

Index entries

PCRprimer designprimer internal stability

Copyright information

© Humana Press Inc. 1995

Authors and Affiliations

  • Wojciech Rychlik
    • 1
  1. 1.National Biosciences Inc.Plymouth