Hieff qpcr sybr green master mix high rox

Twenty-one ESCC tissues and adjacent tissues from ESCC patients were obtained from July 2020 to June 2021 in Fujian Cancer Hospital. The clinical data for qRT-PCR of patients with esophageal squamous cell carcinoma in our institution was shown in Supplementary Table S2. qRT-PCR was applied to verify the expression of the target SPOCD1 using twenty-one pairs of ECA and adjacent normoal esophageal tissues. The primers of SPOCD1 were listed in Table 1, purchased from BioSune (Shanghai, China). An RTⅢ All-in-One Mix with dsDNase (Monad Biotech Co., Ltd, Shanghai, China) was used to synthesize cDNA from 1 µg of total RNA. The qRT-PCR analyses were conducted on the StepOnePlus Real-Time PCR System (Applied Biosystems, Thermo Fisher Scientific Co., Ltd, US) by the Hieff ®qPCR SYBR^® Green Master Mix, High Rox (Yeasen, Biotechnology Co., Ltd, Shanghai, China). The reaction was: 95°C for 10 min, then 41 cycles of 95°C for 15 s and 60°C for 1 min, last 95°C for 15 s. The reference gene was GAPDH, and the relative levels of gene expression were calculated by the 2^−ΔΔCt method.

AURKA expression in 21 NPC and 17 normal tissues was verified through qRT-PCR. The clinical data for qRT-PCR of patients with NPC in our institution was shown in Supplementary Table S1. The primers of AURKA are listed in Supplementary Table S2 and were synthesized by BioSune (Shanghai, China). The internal reference gene was 18S-rRNA. We used RTIII All-in-One Mix and dsDNase (Monad Biotech Co., Ltd., Shanghai, China) to reverse-transcribe 1 µg of total RNA into cDNA. The qRT-PCR validation was performed on the StepOnePlus Real-Time PCR System (Applied Biosystems, Thermo Fisher Scientific Co., Ltd., US) using Hieff^® qPCR SYBR^® Green Master Mix, High Rox (Yeasen, Biotechnology Co. Ltd., Shanghai, China). The reaction system was: 95 °C for 10 min, then 41 cycles of 95°C for 15 s and 60°C for 1 min, last 95°C for 15 s.