GSM6755748: Hela-KO-ZKSCAN3-3; Homo sapiens; RNA-Seq
Total cellular RNA was extracted from cultured cells using RNAiso Plus (Takara, 9108, Japan) according to the manufacturer's protocol. The RNA concentra-tion and purity were quantified using NanoDrop 2000 ultrafine spectrophotometer (Thermo Scientific, Massachusetts, America). And then the genome DNA was elimi-nated with the RNase-free DNase H. The RNA was then reverse-transcribed with 5× FastKing-RT SuperMix kit (TIANGEN, KR118–01, Beijing, China) according to the manufacturer's protocols. The reverse-transcribed cDNA was used as template for qPCR using Genious 2X SYBR Green Fast qPCR Mix (ABclonal, RK21204) according to the manufacturer's protocol. LightCycler 480 II real-time fluorescence quantitative PCR instrument was used for PCR reaction. The primer sequences used for amplifying the target fragments and GAPDH as internal control were listed in Table1. Gene's rela-tive expression levels were calculated by the 2-ΔΔCt (threshold cycle) method.