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PLoS One ; 13(1): e0189188, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29304124

RESUMO

PCR amplification bias is a well-known problem in metagenomic analysis of arthropod communities. In contrast, variation of DNA degradation rates is a largely neglected source of bias. Differential degradation of DNA molecules could cause underrepresentation of taxa in a community sequencing sample. Arthropods are often collected by passive sampling devices, like malaise traps. Specimens in such a trap are exposed to varying periods of suboptimal storage and possibly different rates of DNA degradation. Degradation bias could thus be a significant issue, skewing diversity estimates. Here, we estimate the effect of differential DNA degradation on the recovery of community diversity of Hawaiian arthropods and their associated microbiota. We use a simple DNA size selection protocol to test for degradation bias in mock communities, as well as passively collected samples from actual Malaise traps. We compare the effect of DNA degradation to that of varying PCR conditions, including primer choice, annealing temperature and cycle number. Our results show that DNA degradation does indeed bias community analyses. However, the effect of this bias is of minor importance compared to that induced by changes in PCR conditions. Analyses of the macro and microbiome from passively collected arthropod samples are thus well worth pursuing.


Assuntos
Artrópodes/genética , Artrópodes/microbiologia , Código de Barras de DNA Taxonômico/métodos , DNA/análise , DNA/genética , Microbiota/genética , Animais , Artrópodes/classificação , Biodiversidade , Código de Barras de DNA Taxonômico/estatística & dados numéricos , Primers do DNA , Ecossistema , Havaí , Metagenoma , Metagenômica/métodos , Metagenômica/estatística & dados numéricos , Reação em Cadeia da Polimerase/métodos , Viés de Seleção
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