Emulsion PCR made easy

Biotechniques. 2020 Jul;69(1):421-426. doi: 10.2144/btn-2019-0161. Epub 2020 Apr 27.

Abstract

Emulsion PCR (ePCR) is an important technique that permits amplification of DNA molecules in physically separated picoliter-volume water-in-oil droplets, and thus avoids formation of unproductive chimeras and other artifacts between similar DNA sequences. However, the recovery of ePCR products involves repeated extraction with hazardous organic solvents followed by purification using silica-based columns, making the overall process cumbersome. In this benchmark, we have described a quick ePCR extraction protocol for the purification of ePCR products, which directly employs silica-based DNA purification columns; products purified using this method have been found to be compatible with gene cloning and next-generation sequencing applications. The method described here makes ePCR easy, safe and within the reach of every laboratory.

Keywords: aptamer; chimerization; emulsion PCR; extraction; next-generation sequencing; silica column purification.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • DNA / chemistry
  • Emulsions / chemistry*
  • High-Throughput Nucleotide Sequencing / methods
  • Polymerase Chain Reaction / methods*
  • Silicon Dioxide / chemistry

Substances

  • Emulsions
  • Silicon Dioxide
  • DNA