Optimisation of PCR Reactions Using Primer Chessboarding

J Clin Virol. 2003 Apr;26(3):369-73. doi: 10.1016/s1386-6532(03)00006-4.

Abstract

In-house polymerase chain reaction (PCR) assays are now an integral part of the work of most diagnostic microbiological laboratories. Despite the availability of commercial reagent 'master-mixes' of some PCR reagents, the optimisation of primers still poses a significant problem. Here, we describe a simple method to assess the concentration of primer needed in single round, multiplex, nested and 'real-time' PCR procedures.

MeSH terms

  • Artifacts
  • Computer Systems
  • DNA Primers / chemistry*
  • DNA, Complementary / genetics
  • Dose-Response Relationship, Drug
  • Electrophoresis, Agar Gel
  • Osmolar Concentration
  • Polymerase Chain Reaction / methods*

Substances

  • DNA Primers
  • DNA, Complementary