Comparing 16S rDNA amplicon sequencing and hybridization capture for pea aphid microbiota diversity analysis

BMC Res Notes. 2018 Jul 11;11(1):461. doi: 10.1186/s13104-018-3559-3.

Abstract

Objective: Targeted sequencing of 16S rDNA amplicons is routinely used for microbial community profiling but this method suffers several limitations such as bias affinity of universal primers and short read size. Gene capture by hybridization represents a promising alternative. Here we used a metagenomic extract from the pea aphid Acyrthosiphon pisum to compare the performances of two widely used PCR primer pairs with DNA capture, based on solution hybrid selection.

Results: All methods produced an exhaustive description of the 8 bacterial taxa known to be present in this sample. In addition, the methods yielded similar quantitative results, with the number of reads strongly correlating with quantitative PCR controls. Both methods can thus be considered as qualitatively and quantitatively robust on such a sample with low microbial complexity.

Keywords: 16S; Amplicon sequencing; Hybridization capture; Microbiota; Pea aphid.

MeSH terms

  • Animals
  • Aphids / microbiology*
  • DNA, Ribosomal / genetics*
  • High-Throughput Nucleotide Sequencing
  • Microbiota / genetics*
  • Pisum sativum
  • RNA, Ribosomal, 16S
  • Sequence Analysis, DNA

Substances

  • DNA, Ribosomal
  • RNA, Ribosomal, 16S