GSM7871922: Mouse, Db 7; Mus musculus; RNA-Seq
Total RNA was isolated using an RNeasy mini kit (Qiagen, Germany). Strand-specific libraries were prepared using the TruSeq Stranded Total RNA Sample Preparation kit (Illumina, USA), following the manufacturer's instructions. Briefly, mRNA was enriched with oligo(dT) beads. Following purification, the mRNA is fragmented into small pieces using divalent cations under 94℃ for 8 min. The cleaved RNA fragments are copied into first-strand cDNA using reverse transcriptase and random primers. This is followed by second-strand cDNA synthesis using DNA Polymerase I and RNase H. These cDNA fragments then go through an end repair process, adding a single'A'base, and then ligating the adapters. The products are then purified and enriched with PCR to create the final cDNA library.