In brief, RNA was extracted from cell lines using RNA-Quick Purification Kit (YiShan Biotech). The HiScript II Q RT SuperMix for quantitative polymerase Chain Reaction (Vazyme Biotech) was used to generate the complementary DNA. Quantitative reverse transcription polymerase chain reaction was then conducted using the ABI 7500 FAST RT-PCR system (Applied Biosystems), and Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was set as the internal reference. Relative quantitation was calculated using the ΔΔCt method. The primers we used are presented in Supplementary Table 1.
Hiscript 2 q rt supermix for quantitative
Manufactured by Vazyme
Sourced in China
HiScript II Q RT SuperMix for quantitative is a reagent formulation for reverse transcription and real-time quantitative PCR (RT-qPCR). It is designed to enable efficient and sensitive detection of target RNA sequences.
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2 protocols using hiscript 2 q rt supermix for quantitative
1
Quantitative Analysis of Gene Expression
2
Gene Expression Analysis by qRT-PCR
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Total RNA was extracted using TRIzol reagent (Thermo Fisher Scientific, Waltham, MA, USA) according to the manufacturer’s protocol. cDNA was synthesized using HiScript II Q RT SuperMix for quantitative PCR (qPCR) (plus genomic DNA [gDNA] wiper) (Vazyme Biotech Co., Ltd., Nanjing, China). Quantitative real-time PCR (qRT-PCR) was performed using MonAmp SYBR green qPCR mix (Monad Biotech Co., Ltd., Wuhan, China) on a QuantStudio 3 real-time PCR system (Thermo Fisher Scientific) following the manufacturer’s instructions. The abundance of individual mRNA transcripts in each sample was assayed in triplicate and normalized to the GAPDH mRNA level using the 2–ΔΔCT method. The primers used for qRT-PCR are listed in Table 1 .
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