Fecal Metagenomic Analysis of PEG-Treated Mice

Feces were collected from mice administered PEG for two or four weeks. Feces were immediately frozen in liquid nitrogen and stored in a deep freezer (−80 °C), as storage at room temperature for more than 15 min or in a domestic freezer for more than three days can impact the composition of bacterial taxa in meta 16S analysis [19 (link)]. DNA was extracted from a pellet of feces (≤50 mg) using the ZymoBIOMICS DNA Miniprep kit (Zymo Research, Irvine, CA, USA) following the manufacturer’s protocol. Bead beating was performed for 20 min using a Disruptor Genie instrument (Scientific Industries, Bohemia, NY, USA) at maximum speed.

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Ishibashi R., Matsuhisa R., Nomoto M., Chudan S., Nishikawa M., Tabuchi Y., Ikushiro S., Nagai Y, & Furusawa Y. (2023). Effect of Oral Administration of Polyethylene Glycol 400 on Gut Microbiota Composition and Diet-Induced Obesity in Mice. Microorganisms, 11(8), 1882.

Publication 2023

ZymoBIOMICS DNA Miniprep kit Disruptor Genie instrument

Bacterial Feces Frozen Genie Mice Nitrogen

Corresponding Organization :

Other organizations : Toyama Prefectural University, University of Toyama

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Variable analysis

independent variables

Duration of PEG administration (2 weeks or 4 weeks)

dependent variables

Composition of bacterial taxa in meta 16S analysis

control variables

Immediate freezing of feces in liquid nitrogen
Storage of feces in deep freezer (-80 °C)
Extraction of DNA from a pellet of feces (≤50 mg) using the ZymoBIOMICS DNA Miniprep kit
Bead beating for 20 min using a Disruptor Genie instrument at maximum speed

controls

No positive or negative controls were explicitly mentioned.

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