GSM9045508: Tibialis anterior muscle, septic mouse, Day 1; mouse metagenome; OTHER
Total RNA was extracted from tibialis anterior muscle, blood, liver, and spleen obtained from septic mice and cecal slurry stock using Isogen (Cat# 311-02501, Nippon Gene, Tokyo, Japan) according to the manufacturer's instructions, followed by reverse transcription to cDNA. Reverse transcription was performed using 1 μg of total RNA, random primers (Cat# 3801, Takara Bio, Kusatsu, Japan), RNase inhibitors (Cat# 10777019, Thermo Fisher Scientific), a deoxynucleotide mix (Cat# 639125, Takara Bio), and Moloney murine leukemia virus reverse transcriptase (Cat# 28025013, Thermo Fisher Scientific). Sequencing libraries were prepared by 2-step tailed PCR method targeting the V3/V4 region of the bacterial 16s rRNA gene, following the 16S metagenomic sequencing library preparation protocol for the Illumina MiSeq system. The libraries were sequenced on a MiSeq (Illumina) with a 300-base pair-end read setting.