Sybr green pcr master mix system

RT-qPCR was used to further verify the mRNA expression of the candidate hub genes in TNBC tissues and adjacent tissues. Total RNA from TNBC patients’ tissues was isolated by TRIzol reagent (Invitrogen, Carlsbad, CA, USA). Total RNA quantity was evaluated by a NanoDrop ND-1000 spectrophotometer (Thermo Fisher Scientific, Waltham, MA, USA). RNA was reverse transcribed into cDNA according to the instructions of the Takara kit (Takara Bio Inc., Japan). RT-qPCR reactions were performed using the SYBR Green PCR Master Mix System (Tiangen Biotech, Beijing, China). GAPDH was used as a control to compare the relative expression of NUF2, FAM83D and CENPH mRNA in 14 pairs of triple negative breast cancer paired tissues. Three replicate holes were performed for target genes in the RT-qPCR experiment, and the primer sequences are shown in Table 1. The primers of the target genes and the internal reference gene were synthesized by Sangon Biotech (Shanghai) Co., Ltd.

The RRM2 mRNA expression was detected using reverse transcription quantitative PCR (RT-qPCR) in 18 pairs of LUSC tissues and 12 pairs of LUAD tissues. Total RNA was isolated from NSCLC patient tissue samples using TRIzol reagent (Invitrogen, Carlsbad, CA, USA). Quantity of RNA was determined using a NanoDrop ND-1000 spectrophotometer (Thermo Fisher Scientific, Waltham, MA, USA). Reverse transcription of RNA into cDNA was performed according to the instructions of the Takara kit (Takara Bio Inc., Japan). The SYBR Green PCR Master Mix System (Tiangen Biotech, Beijing, China) was used for RT-qPCR reactions. GAPDH was used as a control for comparison of relative expression of RRM2 mRNA. In the RT-qPCR experiment, three replicate wells were performed. The primer sequences are shown in Table 1, and the primers were supplied by Sangon Biotech (Shanghai) Co., Ltd.