GSM8389841: Wild-type SK36 Rep3; Streptococcus sanguinis; RNA-Seq
SSA_0451 mutant and S. sanguinisSK36 wildtype were cultured in BHI medium overnight,  diluted into fresh BHI medium, and grown for another 4 hours under microaerobic conditions at 37 °C. Samples were collected and treated with RNA protect bacteria reagent (Qiagen, Valencia, CA) for 5 min to stabilize RNA and stored at −80 °C. Cells were lysed by mechanical disruption using FastPrep lysing matrix B (Qbiogene, Irvine, CA). Total RNA was treated with DNase I (Qiagen) and prepared using RNA easy mini kits (Qiagen) according to the manufacturer's instructions.  Total RNA quantity and integrity were determined using a Bioanalyzer (Agilent). All samples passed quality control assessment with RNA Integrity Numbers (RIN) above 8. rRNA depletion was carried out from 100ng of total RNA with NEBNext bacterial rRNA depletion kit (NEB#e7850). NEBNext Ultra II Directional RNA Library Prep Kit for Illumina (New England BioLabs) was used for the following RNA-seq library preparation according to the manufacturer's protocol.