GSM9407361: Control_3 (for ASK1-MKI); Mus musculus; RNA-Seq
Total RNA extraction was conducted by TRlzol Reagent. RNA detect concentration using qubit (Thermo Fisher Scientific, Q33226). RNA purity was checked using the NanoDrop PR-1(Thermo Fisher Scientific,CHEM-PR1-KIT). RNA integrity was assessed using the Qsep1 system (Qsep1 Technologies, CA, China). Messenger RNA was purified from total RNA using attached magnetic beads. After fragmentation, the first strand cDNA was synthesized using primers. Then the second strand cDNA was synthesized using dUTP, instead of dTTP. The directional library was ready after end repair, 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.