GSM8415502: WPMY-1, shvector-1,RNA-seq; Homo sapiens; RNA-Seq
Total RNA was extracted using Trizol reagent Using the structural feature that most eukaryotic mRNAs have a poly(A) tail, mRNAs with poly(A) tails are enriched using Oligo(dT) magnetic beads. Subsequently, the obtained mRNA is randomly fragmented using divalent cations. Using the fragmented mRNA as a template and random oligonucleotides as primers, the first strand of cDNA is synthesized in a 1st Strand Enzyme reverse transcriptase system. Then, 2nd Strand Enzyme and dNTPs are used to synthesize the second strand of cDNA. After purification, the double-stranded cDNA undergoes end repair, A-tailing, and sequencing adapter ligation. cDNA fragments of around 200-300bp are selected, PCR amplified, and the PCR products are purified again, ultimately obtaining the library.