GSM6909158: ATCC49619, normal [B-R1; Streptococcus pneumoniae; RNA-Seq
RNA samples were isolated by TRizol reagent The mRNA was enriched by removing rRNA using the Ribo-zero kit. mRNA was subsequently broken into short fragments by adding fragmentation buffer, and one-stranded cDNA was synthesized using six-base random primers (random hexamers) with mRNA as the template, followed by the addition of buffer, dNTPs (dTTP in dNTP was replaced by dUTP) and DNA polymerase I and RNase H. The double-stranded cDNA was then purified with AMPure XP beads, followed by degradation of the second strand of cDNA containing U with USER enzyme. The purified double-stranded cDNA was first end-repaired, A-tailed, and sequencing junction ligated, and then fragment size selection was performed with AMPure XP beads. Finally, PCR amplification was performed and the PCR products were purified with AMPure XP beads to obtain the final library.