Total RNA was extracted from PC cells or tissues with TRIzol for quantitative analysis. RNA was first reverse transcribed to cDNA with Vazyme reverse transcriptase at 42°C for 45 minutes following the manufacturer’s protocol, and PCR was then performed using SYBR Green qPCR Master Mix (GenePharma). Human ACTB was used as the housekeeping gene for mRNA expression. After 40 cycles of denaturation, annealing, and extension, the relative RNA levels were determined with the 2^-ΔΔCT method relative to the control. All the primers are listed in S1 Table .
Sybr green qpcr master mix
Manufactured by GenePharma
Sourced in China
SYBR Green qPCR Master Mix is a ready-to-use solution for quantitative real-time PCR (qPCR) analysis. It contains all the necessary components, including SYBR Green I dye, DNA polymerase, buffer, and dNTPs, for efficient and sensitive detection of target DNA sequences.
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Lab products found in correlation
3 protocols using sybr green qpcr master mix
1
Quantitative Analysis of mRNA Levels
2
Quantifying mRNA Expression by qRT-PCR
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Total RNA was extracted with TRIzol from PC cells and tissues. All primers used for qRT‐PCR were designed and blasted in the National Center for Biotechnology Information database. RNA was reverse transcribed to cDNA and PCR was then performed using SYBR Green qPCR Master Mix (GenePharma, China) in triplicate. Human β‐actin was used as the internal control for mRNA expression. The primer sequences are shown in Table S1 .
3
Quantifying PC Cell Proliferation
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Total RNA was extracted with TRIzol from PC cells and tissues. All primers used for qRT-PCR were designed and blasted in the National Center for Biotechnology Information database. RNA was reverse transcribed to cDNA with Vazyme reverse transcriptase following the protocol, and PCR was performed using SYBR Green qPCR Master Mix (GenePharma, China) in an ABI CFX Connect real-time PCR system (Bio-Rad, Hercules, USA) in triplicate. Human β-actin was used as the internal control for mRNA expression. The primer sequences are shown in Supplementary table .
CCK-8 proliferation assay CCK-8 assay was used to detect the proliferation ability of PC cells. Transfected cells were seeded in 96well plates at a concentration of 5*10 3 cells per well. After cultured for 0, 24, 48, 72 and 96 hours under the same conditions, PC cells were incubated with diluted CCK-8 reagent (Dojindo Laboratories, Japan) following the instructions. Then, the absorbance was measured in a microplate reader at a wavelength of 450 nm.
CCK-8 proliferation assay CCK-8 assay was used to detect the proliferation ability of PC cells. Transfected cells were seeded in 96well plates at a concentration of 5*10 3 cells per well. After cultured for 0, 24, 48, 72 and 96 hours under the same conditions, PC cells were incubated with diluted CCK-8 reagent (Dojindo Laboratories, Japan) following the instructions. Then, the absorbance was measured in a microplate reader at a wavelength of 450 nm.
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