Total RNA was extracted from cells using RNA isolater (Vazyme, Nanjing, China). Complementary DNA (cDNA) for mRNA or miRNA was then synthesized with HiScript RT SuperMix (Cat # R223-01, Vazyme). MiRNA/U6 snRNA (small nuclear RNA) RT primer mix and PCR specific primer set (GenePharma, China) were used to reversely synthesize cDNA for miRNAs, detect and quantify miRNAs expression, respectively. qRT-PCR was performed with qPCR SYBR® Green Master Mix (Vazyme) on Real-Time PCR system (Applied Biosystems, USA). U6 snRNA or GAPDH was used as an endogenous control for miRNA or mRNA, respectively. 2−∆∆Ct method was carried out to measure the relative expression levels of mRNA or miRNA. The primer sequences were mentioned in our previous study [23 (link)].
Pcr specific primer set
Manufactured by GenePharma
Sourced in China
The PCR specific primer set is a laboratory equipment designed for use in polymerase chain reaction (PCR) experiments. It consists of a pair of short DNA sequences that are complementary to specific regions of the target DNA, allowing for the selective amplification of a particular genetic sequence.
Automatically generated - may contain errors
Lab products found in correlation
Rna isolater, by Vazyme (1 mentions)
Rt primer mix, by GenePharma (1 mentions)
Real time pcr system, by Thermo Fisher Scientific (1 mentions)
Sybr green qpcr master mix, by Vazyme (1 mentions)
Abi prism 7500 sequence detector, by Thermo Fisher Scientific (1 mentions)
M mlv, by Vazyme (1 mentions)
Trizol, by Transgene (1 mentions)
Sybr green master mix, by Vazyme (1 mentions)
2 protocols using pcr specific primer set
1
Quantification of mRNA and miRNA Expression
2
Gene Expression Analysis by qRT-PCR
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Total RNA was extracted using Trizol (TransGen Biotech, China). The first-strand cDNA was reversely transcribed with M-MLV (Vazyme Biotech, China) following the standard protocols. qRT-PCR was carried out to determine mRNA expression with SYBR Green master mix (Vazyme Biotech, China) on an ABI Prism 7500 Sequence Detector (Applied Biosystems, USA). MiRNA/U6 snRNA RT primer mix and PCR specific primer set (GenePharma, China) were used to detect and quantify miRNA expression. Primer sequences of P-gp were described in our previous study [26 (link)]. Other Primer sequences used were mentioned in Supplementary Table 4. mRNA and miRNA levels were normalized to GAPDH or U6 sRNA, respectively. The relative expression was calculated using 2−∆∆Ct method.
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