|Number of samples:||48|
|Release date:||Dec 3 2015|
|Last update date:||Mar 3 2016|
|Dataset link||Gut Microbiota Orchestrates Energy Homeostasis during Cold [16S rRNA gene V4-region miSeq]|
C57BL/6J mice were put for 31 day on cold (6C) or room temperature. Fresh feces and cecum samples were collected, immediately frozen and stored. Bacterial DNA content was extracted using QIAamp Fast DNA stool Mini Kit (Qiagen). Bacterial DNA was PCR amplified with barcoded universal bacterial primers targeting variable regionV4 of 16SrRNA gene. Samples were pooled and sequenced with Ilumina MiSeq platform. Using QIIME and custom scripts, sequences were quality filtered and demultiplexed using exact matches to the supplied DNA barcodes. Resulting sequences were then searched against the Greengenes reference database of 16S rRNA gene sequences, clustered at 97% by uclust. The longest sequence from each Operation Taxonomic Unit (OTU) thus formed was then considered as the OTU representative sequence, and assigned taxonomic classification via Mothur's Bayesian classifier, trained against the Greengenes database clustered at 98%. Each listed sample is indivudual biological replicate.