GSM8966370: S. mutans 0.025uM HP-29, rep 3; Streptococcus mutans UA159; RNA-Seq
Harvested cells were resuspended in 1mL of RNA Protect Bacteria Reagent (Qiagen). Total RNA was isolated from homogenized S. mutans cell lysates by acid-phenol:chloroform extractions.  The nucleic acid was subjected to DNase I digestion (Ambion).   The RNA was then purified using the Qiagen RNeasy colum purification kit, including an additional on-column DNase digest.  Total RNA was subjected to two rounds of mRNA enrichment using the MICROBExpress Bacterial mRNA Purification kit (ThermoFisher). rRNA depletion was performed with the Ribo-Zero Plus Microbiome kit (Illumina) to generate RNA libraries. cDNA librariers were geneated by SeqCenter using an Illumina TrueSeq library prep kit. All sequencing and subsequent steps were performed by SeqCenter.