title: GSM6133676: wt continentalic acid rep1; Streptococcus mutans; RNA-Seq
library_construction_protocol: The cells were harvested, resuspended in 1 mL of RNAprotect Bacteria Reagent (Qiagen, Venlo, Netherlands), and maintained at room temperature for 10 min. The cells were resuspended in 50 mM Tris-HCl buffer containing 10 mM EDTA and 0.4% SDS and then mechanically disrupted using Bead Beater 16 (Biospec Products Inc., Bartlesville, OK). Total RNA was isolated using an RNeasy Mini Kit (Qiagen, Valencia, CA) and the RNA concentration was measured using a nanodrop (NanoDropTM 2000 Spectrophotometer, Thermo Fisher Scientific, Waltham, MA). Prior to the sequencing library construction, the quality and integrity of the extracted RNA were checked using a 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA) and the quantity was measured by quantitative polymerase chain reaction (qPCR) as per the Illumina qPCR Quantification Protocol Guide. One microliter of total RNA was used for the construction of cDNA libraries. Ribosomal RNA was removed by Ribo-ZeroTM Removal Kits (Bacteria) and then converted to cDNA using TruSeq RNA Library Prep Kit v2 (Illumina, San Diego, CA). Paired-end sequencing of the constructed cDNA libraries was performed by Illumina Novaseq.