Total RNA was extracted from cultured cells using TRIzol® reagent (Invitrogen, NY). Single-stranded cDNA was generated from total RNA, using M-MLV reverse transcriptase and oligo (dT) 12–18 primers (Takara, Japan). The real-time quantitative PCR reaction was performed with the SYBR green detection system (Bio SYBR Green Master Mix, Takara, Japan). GAPDH served as an endogenous control. All cycle threshold (ct) values were determined in real time using CFX96™ Real-Time PCR Detection (Bio-rad, CA). The primer pairs for each target gene is listed in Supplementary Table 2 .
Sybr green detection system bio sybr green master mix
Manufactured by Takara Bio
Sourced in Japan
The SYBR green detection system (Bio SYBR Green Master Mix) is a reagent designed for real-time PCR applications. It contains SYBR Green I dye, which binds to double-stranded DNA and emits fluorescence upon binding, allowing for the quantification of DNA amplification during the PCR process.
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Lab products found in correlation
2 protocols using sybr green detection system bio sybr green master mix
1
Quantitative Gene Expression Analysis
2
Quantitative Analysis of Rap2B Expression
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Total RNA was extracted from HCC tissues and cells using TRIzol reagent (Invitrogen) and reverse transcribed into cDNA using a PrimeScript™ 1st Strand cDNA Synthesis kit (Takara, Dalian, China). qRT-PCR was performed with the SYBR Green Detection System (Bio SYBR Green Master Mix, Takara, Japan). The primer sequences used for PCR amplification were Rap2B, 5′-CTGCCCCTTCATGGAGACA-3′ (forward) and 5′-TGCGAATAGCTCATCCACTGA-3′ (reverse); β-actin was used as an internal standard, and the primers were as follows: 5′-GTCCACCGCAAATGCTTCTA-3′ (forward) and 5′-TGCTGTCACCTTCACCGTTC-3′ (reverse). The band intensities of amplification products were measured by a densitometer, and the results were normalized with β-actin.
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