Polymerase chain reaction with nearby primers

Anal Biochem. 2017 Feb 1:518:126-133. doi: 10.1016/j.ab.2016.11.017. Epub 2016 Nov 28.

Abstract

DNA analysis of biological specimens containing degraded nucleic acids such as mortal remains, archaeological artefacts, forensic samples etc. has gained more attention in recent years. DNA extracted from these samples is often inapplicable for conventional polymerase chain reaction (PCR), so for its amplification the nearby primers are commonly used. Here we report the data that clarify the features of PCR with nearby and abutting primers. We have shown that the proximity of primers leads to significant reduction of the reaction time and ensures the successful performance of DNA amplification even in the presence of PCR inhibitors. The PCR with abutting primers is usually characterized by the absence of nonspecific amplification products that causes extreme sensitivity with limit of detection on single copy level. The feasibility of PCR with abutting primers was demonstrated on species identification of 100 years old rotten wood.

Keywords: Abutting primers; Amplification; Degraded DNA; Nearby primers; PCR sensitivity; PCR specificity; Polymerase chain reaction.

MeSH terms

  • DNA Primers / chemistry*
  • DNA Primers / genetics
  • DNA, Plant / chemistry*
  • DNA, Plant / genetics
  • DNA, Plant / isolation & purification*
  • Plants / chemistry*
  • Plants / genetics
  • Polymerase Chain Reaction / methods*

Substances

  • DNA Primers
  • DNA, Plant