GSM8293927: DMSO3; Mus musculus; RNA-Seq
After extracting total RNA from the samples, enrich eukaryotic mRNA using magnetic beads with Oligo(dT) (for prokaryotes, remove rRNA with a kit before proceeding to the next step). Add fragmentation buffer to break the mRNA into short fragments. Using mRNA as a template, synthesize the first cDNA strand with random hexamers (six-nucleotide random primers). Add buffer, dNTPs, RNase H, and DNA polymerase I to synthesize the second cDNA strand. After purification with a QiaQuick PCR kit and elution with EB buffer, perform end repair and ligate sequencing adapters. Use agarose gel electrophoresis for fragment size selection, followed by PCR amplification. Finally, sequence the constructed library.