AAT Bioquest

What factors affect the specificity of PCR?

Posted March 22, 2022


Factors that affect the specificity of PCR include:

  • Length of the primer
  • Purity of the template DNA
  • Low concentrations of template DNA
  • Purity and sequence of the primer
  • Annealing temperature of the PCR reaction
  • Mg2+ concentration
  • Additives such as dimethyl sulfoxide (DMSO)
  • Amounts of PCR inhibitors in the sample such as excessive proteins, SDS, phenol/ethanol, or hemoglobin
  • Inaccurate sample or reagent pipetting particularly when doing serial dilutions
Additional resources

Enhancing the specificity of polymerase chain reaction by graphene oxide through surface modification: zwitterionic polymer is superior to other polymers with different charges

ROX Reference Dye *50X fluorescence reference solution for PCR reactions*

Polymerase Chain Reaction (PCR)