title: GSM4827052: RheMac_SCNT_Placenta_RNA; Macaca mulatta; RNA-Seq
library_construction_protocol: The mRNA was purified from total RNA using oligo-dT attached magnetic beads. Fragmentation was carried out using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer. Then, the first strand cDNA was synthesized using random hexamer primer and M-MuLV Reverse Transcriptase (NEB). Second strand cDNA synthesis was subsequently performed using DNA Polymerase I and RNase H. Remaining overhangs were converted into blunt ends via exonuclease and polymerase activities. After adenylation of 3' ends of DNA fragments, NEBNext Adaptor with hairpin loop structure were ligated to prepare for hybridization. USER Enzyme (NEB) was used to release the looped adaptors. Then the adaptor-ligated DNA was amplified with Phusion High-Fidelity DNA polymerase and purified for sequencing.