title: GSM5272706: B220+ sorted cells from VavPBcl2/RagC Q119L/+ lymphoma [YCX00166]; Mus musculus; RNA-Seq
library_construction_protocol: B220+ were isolated from mouse lymphoma tumours by immunomagnetic enrichment with CD45R (B220) microbeads (Miltenyi Biotech, #130-049-501). Total RNA was extracted using TRIzol and PicoPure RNA Isolation kit (Arcturus, #12204-01) following the manufacturer's instructions Total RNA samples were processed into cDNA sequencing libraries with the "QuantSeq 3' mRNA-Seq Library Prep Kit (FWD) for Illumina" (Lexogen, Cat.No. 015). 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 are completed by PCR, and sequenced on an Illumina instrument (see below) 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}