qPCR was carried out as described before [15 (link)]. The total RNAs were extracted from GES-1, AGS, HGC-27 and MKN-45 cells using TRIzol reagent (T9108, Takara, Japan). After the removal of genomic DNA, the RNAs were subjected to reverse-transcription by Hifair III 1st Strand cDNA Synthesis SuperMix (11141ES10, Takara) for qPCR. The samples were measured using a qRT-PCR kit (QR0100-1KT, Sigma-Aldrich, USA). The results were calculated on a basis of 2-ΔΔCt. Bio-repeats were implemented in triplicate.
Hifair 3 1st strand cdna synthesis supermix
Manufactured by Takara Bio
Sourced in Japan
The Hifair III 1st Strand cDNA Synthesis SuperMix is a reagent kit designed for the reverse transcription of RNA to complementary DNA (cDNA). It contains all the necessary components for the efficient conversion of RNA to first-strand cDNA, which can then be used for various downstream applications such as PCR, qPCR, or gene expression analysis.
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Lab products found in correlation
2 protocols using hifair 3 1st strand cdna synthesis supermix
1
qPCR Analysis of Gene Expression
2
Quantitative Gene Expression Analysis of EC Cell Lines
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Total RNA was extracted from EC cell lines with TRIzol reagent (TaKaRa, Shiga, Japan), and then Hifair® III 1st Strand cDNA Synthesis SuperMix was used for qPCR, and then total RNA reversed to cDNA. Next, Hieff® qPCR SYBR® Green Master Mix was utilized to amplify the target gene by real-time qPCR and measure its expression level. Further, 18sRNA and the U6 gene were acted as an internal reference gene. The related primer sequences were listed in the Table 1 .
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