GSM9236156: tumor, VHH-OD-PD-1-Fc, rep1; Mus musculus; RNA-Seq
Total RNA was extracted using the RNeasy Plus Mini Kit (Qiagen, 74136). RNA was quantified on NanoDrop and RNA quality was assessed on an Agilent 2100 Bioanalyzer. First, poly(A)+ mRNA was enriched from total RNA using oligo(dT)-coated magnetic beads and fragmented. Reverse transcriptase was then used to synthesize the first and second strands of cDNA. The cDNA fragments underwent end repair and A-tailing at the 3' ends, followed by ligation of sequencing adapters. After purification to remove incompletely ligated adapters and adapter dimers, libraries were PCR-amplified using primers complementary to the adapter sequences. Finally, the sequencing libraries were purified using magnetic beads. RNA-seq (paired end 150 bp) was performed by illumina NovaSeq 6000.