GSM9271875: Terc -/- Sting gt/gt, replicate 4; Mus musculus; RNA-Seq
Total RNA was extracted with TRIzol reagent (ThermoFisher 15596026) and then purified and treated with DNAse using RNeasy Kit (QUIAGEN 74106), following manufacturer´s instructions. Provided total RNA samples (average RIN/RQS 5.6, range 3.2-7.1) were converted into sequencing libraries using the "QuantSeq 3' mRNA-Seq V2 Library Prep Kit (FWD) for Illumina" (Lexogen, Cat.No. 191). Briefly, library generation is initiated by reverse transcription with oligodT priming, followed by a random-primed second strand synthesis. A UMI Second Strand Synthesis module was used, in which random primers featuring a 6 nt long Unique Molecular Identifier tag are used {to be addressed for bias correction at data analysis}. Directional cDNA libraries, stranded in the sense orientation {--library-type fr-secondstrand in TopHat, --stranded=yes in HTSeq, --rna-strandness F in HISAT}, were completed by PCR and sequenced in a single read format on an Illumina instrument.