title: GSM8193689: shzmat2-1-2; Homo sapiens; RNA-Seq
library_construction_protocol: RNA was were extracted by Trizol method 1) By utilizing the structural characteristic that most eukaryotic mRNAs carry a polyA tail, mRNAs with polyA tails  were enriched by Oligo(dT) magnetic beads; 2) The purified mRNAs were cleaved into small fragments with fragmentation buffer at a suitable temperature; 3) First-strand cDNAs were produced by reverse transcription using a random hexamer primer; 4) Second-strand cDNAs are synthesized (strand-specific library: dUTPs were used instead of dTTPs in the  second-strand synthesis to incorporate dUTPs in the second-strand cDNAs, while the high-fidelity DNA  polymerase used in this method could not amplify uracil-containing DNA templates, thus realizing the  strand-specificity), while simultaneously performing end repair and dA-Tailing; 5) Sequencing adapter ligation was performed, followed by DNA magnetic bead purification and fragment selection  after ligation was completed to yield a library with 250-350 bp insert fragments; 6) The ligated products were amplified by PCR and purified again using DNA magnetic beads, where the products  were solubilized with nuclear-free water; 7) After the initial library was constructed, a Qubit fluorescence quantifier was used for concentration detection,  followed by a Qsep400 high-throughput biofragment analyzer for fragment size detection，Finally, the  effective concentration of the library was accurately quantified using Q-PCR;