GSM5144639: New-7-1; Hominidae; RNA-Seq
cells were cultured with 4sU for different time (0h, 1h, 4h, and 8h), and RNA was harvested using Trizol reagent.  Novogene RNA library Prep Kit was used with 1 ug of total RNA for the construction of sequencing libraries. A total amount of 1 µg RNA per sample was used as input material for the RNA sample preparations. Briefly, mRNA was purified from total RNA using poly-T oligo-attached magnetic beads. Fragmentation was carried out using divalent cations under elevated temperature in First Strand Synthesis Reaction Buffer. First strand cDNA was synthesized using random hexamer primer and RNase H. In order to select cDNA fragments of preferentially 100~200 bp in length, the library fragments were purified with AMPure XP system (Beckman Coulter, Beverly, USA). Adapter ligation at 25°C for 10min before PCR . Then PCR was performed with Phusion HighFidelity DNA polymerase, Universal PCR primers and Index (X) Primer. At last, PCR products were purified (AMPure XP system) and library quality was assessed on the Agilent Bioanalyzer 2100 system.