Faststart universal 96 sybr green master rox

Total RNA was prepared using TRIzol Reagent (Invitrogen, Carlsbad, CA, United States) according to the manufacturer’s instructions. The concentration of the total RNA was detected by NanoDrop 2000 (Thermo Scientific^TM). Total RNA (1000 ng) was reverse transcribed into cDNA using qPCR RT Kit (TOYOBO, Japan). Relative expression of target gene to the housekeeping gene GAPDH was determined by qRT-PCR using FastStart Universal 96 SYBR Green Master (ROX) (Roche, Germany). All primer sequence used in this study is listed in Supplementary Table 1. Analysis between the two groups was performed by an unpaired t-test; P < 0.05 was considered statistically significant.

According to the manufacturer’s instructions, the total RNA was extracted by TRIzol reagent (Carlsbad Invitrogen, California, United States). The total RNA concentration was detected using a NanoDrop2000 (Thermo Science^TM). RNA (1,000 ng) was reverse transcribed into cDNA using a qPCR RT Kit (TOYOBO, Japan). FastStart Universal 96 SYBR Green Master (ROX) (Roche, Germany) was used to detect the relative expression of TXNDC12 and housekeeping gene GAPDH by qRT-PCR. The primer sequences used in our procedure were as follows: TXNDC12, 5-GTC​CTG​CTG​ATT​GTG​AAA​ATG​GC-3 and 5-TGATCCATGTCGAG GGTCAAA-3; GAPDH, 5-AAT​CCC​ATC​ACC​ATC​TTC-3 and 5-AGGC TGTTGTCATACTTC-3′. A non-paired t-test was performed between the two groups, and p < 0.05 was considered statistically significant.