Total cellular RNA was isolated with RNA Express Total RNA Kit (M050, NCM Biotech, China) (37 (link)). The RNA was reverse transcribed using the RevertAid First Strand cDNA Synthesis Kit (Thermo). We performed RT-qPCR on the StepOne Plus (Applied Biosystems, United States) by utilizing SYBR Green qPCR Master Mix (2×) (Bimake, United States). The primers are listed in Table 1
Sybr green qpcr master mix 2
Manufactured by Selleck Chemicals
Sourced in United States
SYBR Green qPCR Master Mix (2×) is a ready-to-use solution containing all the necessary components, including SYBR Green I, for quantitative real-time PCR (qPCR) reactions. It is designed to simplify and optimize qPCR experiments.
Automatically generated - may contain errors
4 protocols using sybr green qpcr master mix 2
1
Total RNA Extraction and RT-qPCR Analysis
2
Comprehensive RNA Extraction and RT-qPCR Analysis
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Total cellular RNA was extracted using the RNA Express Total RNA Kit (M050, NCM Biotech, China). For cDNA synthesis, reverse transcription was conducted with the RevertAid First Strand cDNA Synthesis kit (K1622, Thermo Fisher Scientific, United States). Subsequently, RT-qPCR was performed on the StepOne Plus (Applied Biosystems, United States) by utilizing SYBR Green qPCR Master Mix (2×) (Bimake, United States). The primers used for the RT-qPCR were listed in Table 1 .
3
Gene Expression Analysis by RT-qPCR
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Isolation of total cellular RNA was performed with RNA Express Total RNA Kit (M050, NCM Biotech. Suzhou, China) [42 (link)]. RevertAid First Strand cDNA Synthesis Kit (Thermo. Waltham, MA, USA) was used to reverse-transcribe the RNA. RT-qPCR was performed on the StepOne Plus (Applied Biosystems. Waltham, MA, USA) by utilizing SYBR Green qPCR Master Mix (2×) (Bimake. Houston, TX, USA). RT-qPCR was conducted using lyceraldehyde-3-phosphate dehydrogenase (GAPDH) as an internal control gene. A list of primers was given in Table 1 .
4
Gene Expression Analysis by RT-qPCR
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Total RNA was isolated using the Total Extraction Reagent RNA Isolator (Vazyme Biotech Co., Ltd., China). The cDNA was generated using the HiScript® II 1st Strand cDNA Synthesis Kit (Vazyme Biotech Co., Ltd., China). Gene expression was quantified by RT-PCR (qRT-PCR) using standard PCR kits and SYBR Green qPCR Master Mix (2×) (Bimake, Shanghai, China) using a CFX Manager™ Software sequence detection system (Bio-Rad Laboratories, USA). GAPDH was the reference gene. All data were analyzed according to the 2-ΔΔCt relative quantification method. The primer sequences are listed in Table 1
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