title: GSM2399981: micro-g rep. 3; Streptococcus mutans; RNA-Seq
library_construction_protocol: A 10-ml aliquot of each HARV culture was centrifuged at 3,000 RPM, 4°C for 20 minutes and immediately placed on ice. Culture supernatants were decanted and 1 ml RNAlater (Ambion) was added to each cell pellet prior to immediate storage at -80°C. RNA was isolated with the RNeasy Kit (Qiagen) and FASTPREP lysing matrix B tubes (MP Biomedical) using previously-described methods. Each RNA sample was then subjected to a second DNase treatment using the TURBO DNA-free™ Kit (Thermo Fisher Scientific) per the manufacturer’s protocols. Lack of contaminating genomic DNA in each RNA sample was determined using quantitative real-time PCR prior to sending ≥ 5 µg of each RNA sample to Seqwright Genomic Services (Houston, TX), who performed all steps of the RNA-seq workflow, data curation, differential expression and corresponding statistical analysis. Each RNA sample was subjected to an mRNA enrichment step and generation of a library of cDNA template molecules of known strand origin using the TruSeq Total mRNA Sample Preparation Kit (Illumina). Sequencing was performed using the Illumina HiSeq 2500 platform, generating 2 x 100bp read lengths for a total of 2 x 10M reads per sample.
identifiers.namespace: GEO
identifiers.id: GSM2399981