Total RNAs from human LF tissues, mouse LF tissues, and cultured mouse LF cells were isolated using the Trizol reagent (Invitrogen) and reverse transcribed into complementary DNA using the HiScript III RT SuperMix for quantitative PCR (qPCR; R302-01, Vazyme, Nanjing, China), following the manufacturer's description. For miRNA, cDNA and quantitative reverse transcription PCR (qRT-PCR) syntheses were performed using the All-in-One miRNA qRT-PCR Detection Kit (GeneCopoeia, Rockville, MD, USA), following the manufacturer's protocol. Next, qPCR was conducted using ChamQ SYBR qPCR Master Mix (Low ROX Premixed; Q331-02, Vazyme) for mRNA and 2 × All-in-One qPCR Mix (GeneCopoeia) for miRNA based on the LightCycler96 PCR system (Roche, Inc., Switzerland). The mRNA quantification of qPCR was normalised to GAPDH expression, and the miRNA quantification was normalised to U48 expression. The comparative 2−△△CT method was used in the analysis. Table S3 presents a list of RT-qPCR primer sequences.
Hiscript 3 rt supermix for quantitative pcr
Manufactured by Vazyme
Sourced in China
HiScript III RT SuperMix for quantitative PCR is a reverse transcription kit designed for high-efficiency cDNA synthesis. It contains a high-performance reverse transcriptase and optimized buffer components for robust and reliable cDNA generation from RNA templates.
Automatically generated - may contain errors
Lab products found in correlation
All in one qpcr mix, by GeneCopoeia (1 mentions)
Chamq sybr qpcr master mix low rox premixed, by Vazyme (1 mentions)
All in one mirna qrt pcr detection kit, by GeneCopoeia (1 mentions)
Lightcycler 96 pcr system, by Roche (1 mentions)
Trizol reagent, by Thermo Fisher Scientific (1 mentions)
Total rna extraction kit, by Fastagen (1 mentions)
2 protocols using hiscript 3 rt supermix for quantitative pcr
1
Quantitative Analysis of mRNA and miRNA
2
Quantifying Tumor Cell and TAM Gene Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
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The tumor cells and TAMs were harvested and RNA was extracted using the total RNA extraction kit (Fastagen; Cat No. RNAfast200). The cDNA was synthesized from 2 µg purified total RNA using the HiScript III RT SuperMix for quantitative PCR (Vazyme; Cat No. R323-01) according to the manufacturer's instructions. The mRNA expression of EGFR and ILT4 in tumor cells and phenotypic markers of TAMs were determined using specific primers and analyzed using the comparative Ct method and normalized to GAPDH level. All experiments were performed in triplicate. The specific primers used are listed in Table S1 .
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