Locked nucleic acids in PCR primers increase sensitivity and performance

Genomics. 2008 Mar;91(3):301-5. doi: 10.1016/j.ygeno.2007.10.016.

Abstract

The incorporation of locked nucleic acids (LNAs) into oligonucleotide primers has been shown to increase template binding strength and specificity for DNA amplification. Real-time PCR and DNA sequencing have been shown to be significantly enhanced by the use of LNAs. Theoretically, increasing primers' binding strength may also increase the sensitivity of conventional PCR, reducing minimum template requirements. We compared LNA-modified PCR primers with their standard DNA counterparts for amplification sensitivity with template amounts as low as 5 pg. Although the results are highly dependent on the design of the LNA primers, large increases in peak height can be achieved from as little as 75 pg, as well as clearer and more complete profiles. Increased amplification success with lower template amounts may also be seen. Additionally, the use of LNAs can enhance multiplexing. Thus, incorporating LNAs into PCR primers can increase amplification success, sensitivity, and performance under a wide range of conditions.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • DNA Primers / chemistry*
  • DNA Primers / genetics
  • Forensic Genetics / methods
  • Forensic Genetics / statistics & numerical data
  • Humans
  • Oligonucleotides / chemistry*
  • Polymerase Chain Reaction / methods*
  • Polymerase Chain Reaction / statistics & numerical data
  • Sensitivity and Specificity

Substances

  • DNA Primers
  • Oligonucleotides
  • locked nucleic acid