Sybr premix ex taq rnase h kit

Total RNA was isolated from tumor tissues Trizol Reagent (Invitrogen Life Technologies) according to manufacturer’s instructions. Primer Script RT reagent Kit with gDNA Eraser (Takara BIO INC, Kusatsu, Shiga,Japan) was used for reverse transcription of RNA. Real-time quantitative PCR was performed using the SYBR Premix Ex Taq RNAse H+ kit (Takara), and the results were analyzed using the Bio-RAD detection system (Bio-RAD, USA).

Total RNA was isolated using Trizol (Invitrogen) according to the manufacturer’s instructions. RNA was reverse-transcribed using Primer Script RT reagent Kit with gDNA Eraser (Takara BIO INC, Kusatsu, Shiga, Japan). Real-time PCR was performed using the SYBR Premix Ex Taq RNAse H + kit (Takara), and analyzed using the Bio-Rad detection system (Bio-Rad, Hercules, CA, USA). The samples were first incubated 5 min at 95 °C, followed by 40 cycles of 10 s at 95 °C and 30 s at 60 °C. The results were calculated (formula: 2-(Ct catalase-Ct EF1)) and matched to the control samples. The primers were produced by Sangon Biotech (Shanghai, China): catalase (sense primer 5′-ccagaagaaagcggtcaagaa-3′; antisense primer 5′-gagatccggactgcacaaag-3′), EF1 (sense primer 5′-cttcactgctcaggtgat-3′; antisense primer 5′-gccgtgtggcaatccaat-3′).

Total RNA was isolated using the RNA-Quick Purification Kit (ESscience, Shanghai, China) according to the manufacturer’s instructions. RNA was reverse-transcribed using the Primer Script RT reagent Kit with gDNA Eraser (Takara BIO INC, Japan). PCR was performed using the SYBR Premix Ex Taq RNAse H+ kit (Takara BIO INC, Japan) and analyzed using the Bio-Rad detection system (Bio-Rad, Hercules, CA, USA). The samples were first incubated 5 min at 95 °C, followed by 40 cycles of 10 s at 95 °C and 30 s at 60 °C. The results were calculated (formula: 2^{-(Ct target-Ct actin)}) and matched to the control samples. The primers sequences are: human PLA2G2A (F:5′-GAAAGGAAGCCGCACTCAGTT-3′, R:5′-CAGACGTTTGTAGCAACAGTCA-3′); human SREBF1 (F:5′-ACAGTGACTTCCCTGGCCTAT-3′, R:5′-GCATGGACGGGTACATCTTCAA-3′); human ACACA (F:5′-TCACACCTGAAGACCTTAAAGCC-3′, R:5′-AGCCCACACTGCTTGTACTG-3′); human ACACB (F:5′-AGAAGACAAGAAGCAGGCAAAC-3′, R:5′-GTAGACTCACGAGATGAGCCA-3′); human ACLY (F:5′-ATCGGTTCAAGTATGCTCGGG-3′, R:5′-GACCAAGTTTTCCACGACGTT-3′); human ACSS2 (F:5′-AAAGGAGCAACTACCAACATCTG-3′, R:5′-GCTGAACTGACACACTTGGAC-3′); human FASN (F:5′-ACAGCGGGGAATGGGTACT-3′, R:5′-GACTGGTACAACGAGCGGAT-3′); human SCD (F:5′-TTCCTACCTGCAAGTTCTACACC-3′, R:5′-CCGAGCTTTGTAAGAGCGGT-3′); human β-actin (F:5′-AACTCCATCATGAAGTGTGAC-3′, R:5′-GATCCACATCTGCTGGAAGG-3′).