Name: GSM7512447
Instrument: NextSeq 500
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: SINGLE
Construction protocol: Once grown, cells were collected by centrifugation at maximum speed for 10 minutes at 4 degrees Celsius. Media was decanted and pellets frozen overnight a -80 degrees Celsius. The next day, pellets were thawed on ice and 700 microliters of buffer RLT added. Buffered cells are mechanically lysed using a bead beater set the highest setting for 45 five seconds a cycle for three cycles with a 5 minute rest between cycles. During the rest, the cells are stored on ice. Once lysed, a Qiagen RNeasy mini kit was used to collect RNA. Contaminating DNA was removed though RNase-free DNase digestion of at least 10 micrograms of RNA determined by nanodrop. To remove the DNase and concentrate the RNA, a GeneJet RNA isolation and concentration kit was used. Lexogen RiboCop rRNA depletion kit for Bacteria followed by Illumina Truseq Stranded mRNA library prep kit.