GSM6857879: YAA00185; 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 (500 ng) were used. Sample RNA Integrity numbers were 7.2 on average (range 6.7-7.9) when assayed on an Agilent 2100 Bioanalyzer. Sequencing libraries were prepared with the "QuantSeq 3' mRNA-Seq Library Prep Kit (FWD) for Illumina" (Lexogen, Cat.No. 015) by following manufacturer instructions. Library generation is initiated by reverse transcription with oligodT priming, and a second strand synthesis is performed from random primers by a DNA polymerase. Primers from both steps contain Illumina-compatible sequences. Libraries are completed by PCR. This kit generates directional libraries stranded in the sense orientation (the read1 (the only read in single read format) has the sense orientation. cDNA libraries are purified, applied to an Illumina flow cell for cluster generation and sequenced on an Illumina NextSeq 550 (with v2.5 reagent kits) by following manufacturer's protocols.