A Comparison of Three Automated Nucleic Acid Extraction Systems for Human Stool Samples

Automated nucleic acid extractors are useful instruments for the high-throughput processing of bio-samples and are expected to improve research throughput in addition to decreased inter-sample variability inherent to manual processing. We evaluated three commercial nucleic acid extractors Bioer GenePure Pro (Bioer Technology, Hangzhou, China), Maxwell RSC 16 (Promega Corporation, Madison, WI, USA), and KingFisher Apex (ThermoFisher Scientific, Waltham, MA, USA) based on their DNA yield, DNA purity, and 16S rRNA gene amplicon results using both human fecal samples and a mock community (ZymoBIOMICS Microbial Community Standard (Zymo Research Corp., Irvine, CA, USA)). Bead-beating provided incremental yield to effectively lyse and extract DNA from stool samples compared to lysis buffer alone. Differential abundance analysis and comparison of prevalent bacterial species revealed a greater representation of Gram-positive bacteria in samples subjected to mechanical lysis, regardless of sample type. All three commercial extractors had differences in terms of yield, inter-sample variability, and subsequent sequencing readouts, which we subsequently share in the paper and believe are significant considerations for all researchers undertaking human fecal microbiota research.

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Publication Details

Journal
Microorganisms
Published
2024-11-25
DOI
https://doi.org/10.3390/microorganisms12122417
Citations
1
Primary Topic
Gut microbiota and health
Type
article
Field-Weighted Citation Impact
0.23

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article

A Comparison of Three Automated Nucleic Acid Extraction Systems for Human Stool Samples

Wit Thun Kwa, Choon Kiat Sim, Jonathan Lee, Adrian Low
1 citations
Microorganisms
Gut microbiota and health
0.23
article

A Comparison of Three Automated Nucleic Acid Extraction Systems for Human Stool Samples

Wit Thun Kwa, Choon Kiat Sim, Jonathan Lee, Adrian Low
article en
1 citations

Abstract

Automated nucleic acid extractors are useful instruments for the high-throughput processing of bio-samples and are expected to improve research throughput in addition to decreased inter-sample variability inherent to manual processing. We evaluated three commercial nucleic acid extractors Bioer GenePure Pro (Bioer Technology, Hangzhou, China), Maxwell RSC 16 (Promega Corporation, Madison, WI, USA), and KingFisher Apex (ThermoFisher Scientific, Waltham, MA, USA) based on their DNA yield, DNA purity, and 16S rRNA gene amplicon results using both human fecal samples and a mock community (ZymoBIOMICS Microbial Community Standard (Zymo Research Corp., Irvine, CA, USA)). Bead-beating provided incremental yield to effectively lyse and extract DNA from stool samples compared to lysis buffer alone. Differential abundance analysis and comparison of prevalent bacterial species revealed a greater representation of Gram-positive bacteria in samples subjected to mechanical lysis, regardless of sample type. All three commercial extractors had differences in terms of yield, inter-sample variability, and subsequent sequencing readouts, which we subsequently share in the paper and believe are significant considerations for all researchers undertaking human fecal microbiota research.

MicroorganismsVol. 12(12)
National University of Singapore (SG), National University Hospital (SG)
National University of Singapore
Openalex Percentile: Top 41%
Gut microbiota and health
0.23
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