title: GSM6033363: CRC_patient2; Homo sapiens; RNA-Seq
library_construction_protocol: Each sample's total RNA was extracted using TRIzol Reagent (Invitrogen, Carlsbad, CA, USA) according to the manufacturer's instructions. The cDNA library is constructed by enriching mRNA with polyA tail from total RNA by Oli-go (dT) magnetic beads, and then randomly fragmenting the obtained mRNA with diva-lent cations in Fragmentation Buffer. The first strand of cDNA was then synthesized in the M-MuL V reverse transcriptase system using random oligonucleotides as primers and frag-mented mRNA as templates. The second strand of cDNA was synthesized under the DNA polymerase I system using dNTPs from RNaseH-degraded RNA strand. The purified dou-ble-stranded cDNA was end-repaired, A-tailed, and connected to a sequencing adapter. AMPure XP beads were used to screen cDNAs of about 370-420 bp, followed by PCR am-plification and AMPure XP beads were again used to purify the PCR products. At last, we obtained a library.