GSM5008792: BMDMs, Unstimulated, Irg1_KO-1; Mus musculus; RNA-Seq
mRNA was extracted from cell pellets using oligo-dT beads (Invitrogen). Custom oligo-dT primers containing barcode and adaptor-linker sequences (CCTACACGACGCTCTTCCGATCT-XXXXXXXX-T15) were used to for cDNA synthesis. Barcoded first-strand synthesis cDNAs for each sample were pooled together, and the RNA-DNA hybrid was degraded by acid-alkali treatment. A second sequencing adaptor (AGATCGGAAGAGCACACGTCTG) was ligated using T4 ligase (NEB) followed by SPRI bead cleanup (Beckman-Coulter). The libraries were PCR amplified 12 cycles, followed by SPRI bead clean-up prior to sequencing. 3'end DGE single-end demultiplexed