GSM8942100: LU_RAS_KD_D10_AA; Homo sapiens; RNA-Seq
RNA was havested using Rneasy mini plus kit (Qiagen). 200ng of total RNA was used for the construction of sequencing libraries. Sequencing libraries were use Fast RNA-seq Lib Prep Kit V2. RNAwaspurified from total RNA using poly-T oligo-attached magnetic beads. After fragmentation, the first strand cDNA was synthesized using random hexamer primers. Then the second strand cDNA was synthesized using dUTP, instead of dTTP. The directional library 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.