RNA-Seq_PNT_D7 STM_rep1 (CRX1623076)
The VAHTS Total RNA-seq (H/M/R) Library Prep Kit for Illumina (VAHTS, Catalog No. NR603) was utilized for RNA library preparation. Briefly, 1 ug of total RNA was hybridized with the rRNA probe (H/M/R) and treated with RNase H to remove ribosomal RNA. DNase I digestion was subsequently performed to further purify the ribosomal RNA-depleted sample. The RNA was fragmented at 94 degrees Celsius for 8 minutes, followed by the synthesis of first-strand and second-strand cDNA using the provided reagents. The resulting cDNA was purified using VAHTS DNA Clean Beads (VAHTS, Catalog No. NR421), then underwent end repair, dA-tailing, adapter ligation, and digestion of the second strand. After two rounds of purification, the cDNA was PCR-amplified and further purified using VAHTS DNA Clean Beads.