title: GSM5983689: Basal virgin KO [825kBa]; Mus musculus; RNA-Seq
library_construction_protocol: Rna was extracted using the mini Rneasy kit from Quiagen Quantseq 3'UTR; Total RNA samples, 500 ng with an average RIN=6.9 (range: 2.5-8.6) were processed into cDNA sequencing libraries with the "QuantSeq 3' mRNA-Seq Library Prep Kit (FWD) for Illumina" (Lexogen, Cat.No. 015) with a UMI Second Strand Synthesis module. Briefly, library generation is initiated by reverse transcription with oligodT priming, followed by a random-primed second strand synthesis. While primers from both steps contain Illumina-compatible sequences, random primers additionally feature 6 nt long Unique Molecular Identifier (UMI) tags. Libraries are 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, i.e., the read1 (the only read in single read format) has the sense orientation.