Fecal DNA Extraction Protocol

Frozen fecal samples were thawed on ice, 100 mg of each sample was suspended in 4 M guanidium thiocyanate, 100 mM Tris-HCl (pH 9.0), and 40 mM EDTA, and the samples were then beaten with zirconia beads using a FastPrep FP100A instrument (MP Biomedicals, USA). DNA was extracted from the bead-treated suspensions using a Magtration System 12GC and GC series MagDEA DNA 200 (Precision System Science, Japan). DNA concentrations were estimated by spectrophotometry using an ND-1000 instrument (NanDrop Technologies, USA), and the final concentration of the DNA sample was adjusted to 10 ng/µL.

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Takahashi S., Tomita J., Nishioka K., Hisada T, & Nishijima M. (2014). Development of a Prokaryotic Universal Primer for Simultaneous Analysis of Bacteria and Archaea Using Next-Generation Sequencing. PLoS ONE, 9(8), e105592.

Publication 2014

FastPrep FP100A

Edta Fecal Frozen Spectrophotometry Thiocyanate Tris Zirconia

Corresponding Organization :

Other organizations : Okayama Prefecture

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Protocol cited in 30 other protocols

Variable analysis

independent variables

Thawing frozen fecal samples on ice
Suspending 100 mg of each sample in 4 M guanidium thiocyanate, 100 mM Tris-HCl (pH 9.0), and 40 mM EDTA
Beating the samples with zirconia beads using a FastPrep FP100A instrument

dependent variables

DNA concentration estimated by spectrophotometry using an ND-1000 instrument

control variables

Adjusting the final concentration of the DNA sample to 10 ng/μL

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