GSM2058479: Spneumoniae_R6_SECO_t15_rep_2; Streptococcus pneumoniae R6; RNA-Seq
RNA was extracted with Mini Rneasy columns from Quiagen and treated with DNase I 1 µg samples of RNA depleted of ribosomal RNA were chemically fragmented. They were 5′ end-tagged cDNA (equivalent to the 3′ end of the original RNA) by random-primed cDNA synthesis. This was followed by 3′-terminal tagging of the cDNA using the Terminal-Tagging Oligo (TTO) which randomly annealed to the cDNA, and served as template for extension by DNA polymerase. The resulting di-tagged cDNAs were purified using the Ampure beads XP system. Enrichment and barcoding of the purified di-tagged cDNAs was done with 15 cycles of PCR. Library size and quantification by qPCR were determined with a 2100 Bioanalyzer Instrument (Agilent Technologies) and Kapa library quantification kit (Kapa Biosystems), respectively. Library sequencing was performed in a NextSeq 500 platform using a 1 x 75 base run. 29.8-35.47 millions of passing filters reads per sample were obtained.
GEO Accession
GSM2058479