GSM4822331: 2FL_rep2 [2B]; Bifidobacterium longum subsp. infantis; RNA-Seq
Cell pellets stored in RNAlater were centrifuged at 14,000 × g for 3 min and cell pellets were washed with 1 mL cold 1X PBS buffer. Total RNA was extracted using Ambion RNAqeous kit with slight modifications. Accordingly, cells were suspended in 600 μL lysis buffer in beadbeating tubes using a FastPrep 24 bead beader (MP Biomedicals, USA). Total RNA, then,  were subjected to DNase treatment using the Ambion Turbo DNA-free kit. The RNA was evaluated for genomic DNA contamination using qRT-PCR. ribosomal RNA is depleted via RiboMinus kit with Magnetic Bead Clean up Module using the custom Pan-Prokaryote Probe Mix to target bacterial rRNA. Purified mRNA was used asseses for quality and used for RNA-seq library preparation. mRNA enriched library preparation was performed with the NEBNext Ultra II Directional kit following the manufacturer's instructions