GSM9289877: Sample 2, CD22 knockout BMDMs, IL-4-stimulated M2, 48 hours; Mus musculus; RNA-Seq
Rneasy total RNA extraction according to the manufacturer's protocol. RNA integrity was checked with Agilent Technologies 2100 Bioanalyzer. Poly(A) RNA sequencing library was prepared following Illumina's TruSeq-stranded-mRNA sample preparation protocol. Poly(A) tail-containing mRNAs were purified using oligo-(dT) magnetic beads with two rounds of purification. After purification, poly(A) RNA was fragmented using divalent cation buffer in elevated temperature, followed by first strand synthesis, second strand synthesis with dUTP, end repair, 3'adenylation, adapt ligation, UDG treatment, and finally PCR. Quality control analysis and quantification of the sequencing library were performed using Agilent Technologies 2100 Bioanalyzer High Sensitivity DNA Chip. Paired-ended sequencing was performed on Illumina's NovaSeq 6000 sequencing system.