Frozen fecal samples were processed for DNA isolation using the MoBio PowerSoil kit according to the manufacturer’s instructions and sequenced using the Illumina MiSeq platform. Data was processed using the QIIME analysis pipeline (Caporaso et al., 2010 (link)).
Mobio powersoil kit
Manufactured by Illumina
Sourced in United States
The MoBio PowerSoil kit is a DNA extraction and purification system designed to isolate high-quality genomic DNA from a variety of soil and environmental samples. The kit utilizes a bead-beating method to efficiently disrupt cells and release DNA, followed by a series of purification steps to remove inhibitors and contaminants. The extracted DNA can be used for downstream applications such as PCR, sequencing, or other molecular analyses.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using mobio powersoil kit
1
Fecal DNA Isolation and Sequencing
2
Shotgun Metagenomic Sequencing of Gut Microbiome
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The gut microbiome was characterized by shotgun metagenomic sequencing using established methods [34 (link)]. Genomic DNA was extracted using the MO BIO PowerSoil kit and prepared for sequencing with the Nextera XT kit (Illumina, Inc., San Diego, CA, USA). Libraries were sequenced on Illumina HiSeq 2500 using the paired-end 125 bp sequencing protocol. Sequence reads were processed to remove adapter sequences and low-quality reads with Trimmomatic v. 0.33 [35 (link)]. Taxonomic assignments were generated with MetaPhlAn2 [36 (link)]. Gene orthologs were assigned by aligning reads to the KEGG database [37 (link)] using DIAMOND v. 2.0.4 [38 (link)]. Negative (mock purification of DNA-free water) and positive (standard fecal and pond-sediment samples) controls were included in each assay. Samples of each formula (CMF, EHF) were assayed for DNA content, which yielded no detectable bacterial DNA. Shotgun libraries were generated from 1 ng of DNA using the NexteraXT kit (Illumina, Inc.). Libraries were sequenced on the Illumina HiSeq using 2 × 125 bp chemistry in high-output mode.
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