GSM3445735
GSM3445735: del223; Mus musculus; RNA-Seq
Total RNA was extracted with TRIzol reagent (ThermoFisher 15596026) and then purified and treated with DNase using RNeasy Kit (QIAGEN 74106), following manufacturer´s instructions. Total RNA (1µg) was used for the RNA-seq experiment, and the obtained sample RNA Integrity Number was 10 (Agilent 2100 Bioanalyzer). The poly(A)+ fraction was purified and randomly fragmented, converted to double stranded cDNA and processed through subsequent enzymatic treatments of end-repair, dA-tailing, and ligation to adapters with "NEBNext Ultra II Directional RNA Library Prep Kit for Illumina" (NEB, Cat. No. E7760) as recommended by the manufacturer. This kit incorporates dUTP during 2nd strand cDNA synthesis, which implies that only the cDNA strand generated during 1st strand synthesis is eventually sequenced. Adapter-ligated library was completed by PCR with Illumina PE primers. The resulting purified cDNA library was applied to an Illumina flow cell for cluster generation and sequenced on an Illumina NextSeq 500 sequencer, by following manufacturer's protocols.
GEO
GSM3445735
GEO Accession
GSM3445735