GSM1161550: mir217Tg_CD19_DN_R2; Mus musculus; RNA-Seq
Total RNA was extracted with Trizol (Invitrogen). RNA samples were first processed through the "Ovation RNA-seq system" (NuGEN) and then retrotranscribed, and cDNA was amplified by the SPIA isothermal linear amplification procedure. The resulting ds-cDNA was fragmented to a range of 150-200bp in a Covaris S2 shearing instrument cDNA (200ng of per sample) was processed through succeeding enzymatic treatments of end-repair, dA-tailing, and ligation to indexed adapters, according to "TruSeq DNA Sample Preparation Guide" (Illumina). Ligation products were purified with AMPure XP Beads. Adapter-ligated libraries were amplified by PCR with Illumina PE primers for 10 cycles. The resulting purified DNA libraries were applied to an Illumina flow cell for cluster generation, and sequenced on the Illumina Genome Analyzer IIx with SBS TruSeq v5 reagents. Image analysis and per-cycle basecalling were performed with Illumina Real Time Analysis software (RTA1.9)
GEO Accession
GSM1161550