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

BMC Res Notes

Laboratoire de Biométrie et Biologie Evolutive, CNRS, UMR 5558, Université de Lyon, Université Lyon 1, 43 Boulevard du 11 novembre 1918, 69622, Villeurbanne, France.

Published: July 2018

AI Article Synopsis

  • Targeted sequencing of 16S rDNA amplicons is commonly used for profiling microbial communities but has limitations like primer bias and short read lengths.
  • The study compared two PCR primer pairs with a new gene capture method using metagenomic extracts from pea aphids to evaluate their performance.
  • Both methods successfully identified the 8 known bacterial taxa in the sample and showed similar quantitative results, indicating they are reliable for low microbial complexity samples.

Article 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.

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Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6042230PMC
http://dx.doi.org/10.1186/s13104-018-3559-3DOI Listing

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