Blocking primers reduce co-amplification of plant DNA when studying bacterial endophyte communities

J Microbiol Methods. 2015 Oct:117:1-3. doi: 10.1016/j.mimet.2015.07.003. Epub 2015 Jul 6.

Abstract

A blocking primer set based on the technique described by Vestheim and Jarman (2008) was developed to reduce amplification of non-target plant DNA when conducting metagenomic studies on bacterial endophyte communities. Bacterial amplification efficiency was increased 300-fold compared to standard PCR in an Illumina-based study of Sorghastrum nutans leaves.

Keywords: Blocking; Chloroplast; Endophyte; Metagenomics; Mitochondria.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Bacteria / genetics*
  • Bacteria / isolation & purification
  • Chloroplasts / genetics
  • DNA Primers / genetics
  • DNA, Plant / chemistry
  • DNA, Plant / genetics*
  • Endophytes / genetics*
  • Endophytes / isolation & purification
  • Metagenomics / methods*
  • Mitochondria / genetics
  • Plants / genetics
  • Plants / microbiology*

Substances

  • DNA Primers
  • DNA, Plant