GSM6857861: YAA00455; Mus musculus; RNA-Seq
Total RNA was extracted from tissues with TRItidy (Panreac Applichem, #A4051,0200) and using RNeasy kit from Qiagen (#74106) according to manufacturer´s instructions Total RNA samples (500ng; RQS/RIN average 7.1, range 6.5-8.0) were converted into cDNA sequencing libraries with the "QuantSeq 3' mRNA-Seq Library Prep Kit (FWD) for Illumina" (Lexogen, Cat.No. 015). Briefly, library generation is initiated by reverse transcription with oligodT priming, followed by a random-primed second strand synthesis. Primers from both steps contain Illumina-compatible sequences. Libraries were completed by PCR, and sequenced on an Illumina NextSeq 550 (with v2.5 reagent kits) by following manufacturer's protocols. This library preparation kit generates directional libraries stranded in the sense orientation (the read1 (the only read in single read format) has the sense orientation.