Fecal Metagenomic Sequencing Protocol

DNA was isolated from 100 mg of frozen feces using the Qiagen DNeasy PowerSoil Pro Kit (Valencia, CA, USA), and metagenomic sequencing was performed by CosmosID, Inc. (Rockville, MD, USA). As previously described [33 (link)], DNA libraries were prepared using the Illumina Nextera XT preparation kit (San Diego, CA, USA). Library quantity was assessed using a Qubit Fluorometer (Thermo Fisher Scientific, Wilmington, DE, USA) and sequenced on an Illumina HiSeq platform to generate 150-bp paired-end reads.

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Bawaneh A., Wilson A.S., Levi N., Howard-McNatt M.M., Chiba A., Soto-Pantoja D.R, & Cook K.L. (2022). Intestinal Microbiota Influence Doxorubicin Responsiveness in Triple-Negative Breast Cancer. Cancers, 14(19), 4849.

Publication 2022

DNeasy PowerSoil Pro Kit Nextera XT preparation kit Qubit Fluorometer HiSeq platform

Feces Frozen Library Metagenomic

Corresponding Organization : Wake Forest University

Other organizations : Duke University Hospital, Duke Medical Center

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Variable analysis

independent variables

DNA isolation method (Qiagen DNeasy PowerSoil Pro Kit)

dependent variables

Metagenomic sequencing data

control variables

Amount of feces used (100 mg)
Library preparation method (Illumina Nextera XT)
Sequencing platform (Illumina HiSeq)
Sequencing read length (150-bp paired-end)

controls

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