GSM6383286: manL mutant, Glucose, rep 3; Streptococcus sanguinis SK36; RNA-Seq
Bacterial cells were resuspended in a lysis buffer (50 mM Tris-HCl pH 8.0, 10 mM EDTA, and 0.4% SDS) and disrupted by vigorous shaking for 1 min together with an equal volume of acidic phenol: chloroform (1:1, v/v) and similar amount of glass beads. After 10 min of centrifugation at 15,000× g at room temperature, the clarified aqueous layer was removed and processed using an RNeasy mini kit (Qiagen, Germantown, MD) for extraction of total RNA. Genomic DNA contamination was removed in column using an RNase-free DNase I solution (Qiagen). Total RNA is treated with RiboZero Plus rRNA depletion kit to remove rRNA, followed by standard procedure for Illumina cDNA library preparation and sequencing