2.5.2 cDNA synthesis and relative quantitative PCR
Total RNA was retro-transcribed by Evo M-MLV RT Mix Kit on Bio-Rad
T100TM Thermal Cycler. The relative quantitative PCR
was performed with SYBR qPCR Master Mix on Applied Biosystems 7500
Real-time PCR System. Data were analyzed using the 7500 software v2.0.6.
The primers for qRT-PCR of RNAi groups and CRISPR-Cas9 cells are
RNAi-QF/QR and CRISPR-QF/QR (listed in Supplementary Table 1). 18S
ribosomal DNA (rDNA) was regarded as an internal reference for qRT-PCR,
and its primers were listed as 18S-F/R in Supplementary Table 1.