Optimization of subsampling, decontamination, and DNA extraction of difficult peat and silt permafrost samples.
Sci Rep
; 10(1): 14295, 2020 08 31.
Article
em En
| MEDLINE
| ID: mdl-32868827
ABSTRACT
This study aims to act as a methodological guide for contamination monitoring, decontamination, and DNA extraction for peaty and silty permafrost samples with low biomass or difficult to extract DNA. We applied a biological tracer, either only in the field or both in the field and in the lab, via either spraying or painting. Spraying in the field followed by painting in the lab resulted in a uniform layer of the tracer on the core sections. A combination of bleaching, washing, and scraping resulted in complete removal of the tracer leaving sufficient material for DNA extraction, while other widely used decontamination methods did not remove all detectable tracer. In addition, of four widely used commercially available DNA extraction kits, only a modified ZymoBIOMICS DNA Microprep kit was able to acquire PCR amplifiable DNA. Permafrost chemical parameters, age, and soil texture did not have an effect on decontamination efficacy; however, the permafrost type did influence DNA extraction. Based on these findings, we developed recommendations for permafrost researchers to acquire contaminant-free DNA from permafrost with low biomass.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Solo
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DNA
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Sedimentos Geológicos
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Pergelissolo
Idioma:
En
Ano de publicação:
2020
Tipo de documento:
Article