PLoS ONE (Jan 2011)

Decontamination of MDA reagents for single cell whole genome amplification.

  • Tanja Woyke,
  • Alexander Sczyrba,
  • Janey Lee,
  • Christian Rinke,
  • Damon Tighe,
  • Scott Clingenpeel,
  • Rex Malmstrom,
  • Ramunas Stepanauskas,
  • Jan-Fang Cheng

DOI
https://doi.org/10.1371/journal.pone.0026161
Journal volume & issue
Vol. 6, no. 10
p. e26161

Abstract

Read online

Single cell genomics is a powerful and increasingly popular tool for studying the genetic make-up of uncultured microbes. A key challenge for successful single cell sequencing and analysis is the removal of exogenous DNA from whole genome amplification reagents. We found that UV irradiation of the multiple displacement amplification (MDA) reagents, including the Phi29 polymerase and random hexamer primers, effectively eliminates the amplification of contaminating DNA. The methodology is quick, simple, and highly effective, thus significantly improving whole genome amplification from single cells.