Total RNA was extracted using RiboEX (GeneAll, Seoul, Republic of Korea). cDNA was synthesized using a GoScript™ Reverse Transcript kit (Promega, WI, USA), according to the manufacturer’s instructions. SYBR Green PCR Mixture (Toyobo, NY, USA) was used for qRT-PCR analysis. Expression data were acquired using a StepOnePlus™ Real-Time PCR System (Applied Biosystems, CA, USA). Expression levels of each target were calculated using the 2−ΔΔCt method and were presented as relative mRNA expression. The sequences of primers used for qRT-PCR were previously described [37 (link)].
Sybr green pcr mixture
Manufactured by Toyobo
Sourced in United States
SYBR Green PCR Mixture is a ready-to-use solution for real-time polymerase chain reaction (PCR) experiments. It contains SYBR Green I dye, DNA polymerase, dNTPs, and necessary buffers. The SYBR Green I dye binds to double-stranded DNA, allowing the detection and quantification of PCR amplification.
Automatically generated - may contain errors
Lab products found in correlation
Steponeplus real time pcr system, by Thermo Fisher Scientific (1 mentions)
Riboex, by GeneAll (1 mentions)
Goscript reverse transcript kit, by Promega (1 mentions)
M mlv reverse transcriptase, by Thermo Fisher Scientific (1 mentions)
Trizol reagent, by Thermo Fisher Scientific (1 mentions)
Random primer, by Thermo Fisher Scientific (1 mentions)
Icycler cfx96 real time pcr detection system, by Bio-Rad (1 mentions)
2 protocols using sybr green pcr mixture
1
Quantitative RNA Expression Analysis
2
Quantitative RT-PCR for Gene Expression
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RT-qPCR was performed as follows. The total RNA was extracted from cells using a TRIzol reagent (Invitrogen, Carlsbad, CA, USA) according to the manufacturer’s instructions. The cDNA was synthesized with 1 μg of the total RNA using MMLV reverse transcriptase and random primers (Invitrogen). The qPCR was performed using the Icycler CFX96 real-time PCR detection system (Bio-Rad, Hercules, CA, USA) and SYBR Green PCR mixture (Toyobo Co., Ltd., Osaka, Japan). The primers used for qPCR are shown in Supplementary Table S1 . The expression levels were normalized to those of the internal standard GAPDH. The expression level was expressed as the relative change to controls.
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