title: GSM4817408: ML-2-overexpressing empty vector rep1 [ML2C1]; Homo sapiens; RNA-Seq
library_construction_protocol: RNA was extracted using Trizol reagent. RNA libraries were prepared for sequencing using standard Illumina protocols. 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 NEBNext First Strand Synthesis Reaction Buffer (5X). First strand cDNA was synthesized using random hexamer primer and RNase H. Second strand cDNA synthesis was subsequently performed using buffer, dNTPs, DNA polymerase I and RNase H. The library fragments were purified with QiaQuick PCR kits and elution with EB buffer, then terminal repair,A-tailing and adapter added were implemented. The aimed products were retrieved and PCR was performed, then the library was completed.