GSM7566848: ZNF143KO_RNA-seq; Homo sapiens; RNA-Seq
About 1 million cells were used for each RNA-seq experiment. Cells were washed twice using PBS and lysed by 1 mL Trizol (Invitrogen 15596026) for 15 min at room temperature. 0.2 mL chloroform was added and incubated for 2-3 minutes at room temperature. The upper phase was transferred into a new tube after centrifuged at 12,000rpm for 15 min at 4°C, and mixed with 0.5 mL isopropanol. After incubation at room temperature for 10 minutes, the total RNA was precipitated by centrifuging at 12,000 rpm for 10 minutes at 4°C. The precipitated DNA was washed by 75% ethanol to remove salt. Air-dried RNA was dissolved by 100 nuclease-free H2O. The RNA was purified using the RNeasy kit (QIAGEN 75142) to remove residual DNA. 1ug RNA was used for extract mRNA by NEBNext poly(A) mRNA Magnetic Isolation Module (NEB E7490S), and RNA-seq library was constructed using NEBNext®Ultra™II RNA Library Prep Kit. The library was sequenced on the Illumina HiSeq platform.