Dual primer emulsion PCR for next- generation DNA sequencing

Biotechniques. 2010 May;48(5):409-12. doi: 10.2144/000113423.

Abstract

We have developed a highly sensitive single-molecule clonal amplification method called dual primer emulsion PCR (DPePCR) for next-generation DNA sequencing. The approach is similar in concept to standard emulsion PCR; however, in DPePCR both primers are attached to the beads, therefore following PCR amplification, both strands of the PCR products are attached to the beads. The ends of each strand can be freed for analysis by restriction digestion of the bridged PCR fragments, which allows efficient paired-end sequencing of fragment libraries.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • DNA Primers / metabolism*
  • Emulsions
  • Polymerase Chain Reaction / methods*
  • Reproducibility of Results
  • Sequence Analysis, DNA / methods*
  • Streptococcus pyogenes / genetics

Substances

  • DNA Primers
  • Emulsions