title: GSM9567954: Sample_40; Homo sapiens; RNA-Seq
library_construction_protocol: Total RNA was extracted using column-based purification with RNeasy mini kit (Qiagen). One sample of total RNA was shipped to Novogene Europe. RNA quantitation, integrity and purity was evaluated with Agilent 5400 fragment analyzer, before using for the construction of sequencing libraries. The library was checked with Qubit and real-time PCR for quantification and bioanalyzer for size distribution detection Messenger RNA was purified from total RNA using poly-T oligo-attached magnetic beads. After fragmentation, the first strand cDNA was synthesized using random hexamer primers, followed by the second strand cDNA synthesis using either dUTP for directional library or dTTP for non-directional library . For the non-directional library, it was ready after end repair, A-tailing, adapter ligation, size selection, amplification, and purification. For the directional library, it was ready after end repair, A-tailing, adapter ligation, size selection, USER enzyme digestion, amplification, and purification . The library was checked with Qubit and real-time PCR for quantification and bioanalyzer for size distribution detection. Quantified libraries will be pooled and sequenced on Illumina platforms, according to effective library concentration and data amount