After tissue grinding, RNA was extracted with RNAiso Plus (Invitrogen, USA) according to the manufacturer's instructions. cDNA was obtained by reverse transcription using a HisScript® III RT SuperMix for qPCR (+gDNA wiper) kit (Vazyme, China). The qPCR experiment was carried out according to the instructions of the ChamQ Universal SYBR qPCR Master Mix kit (Vazyme, China). The operating instrument used was a Roche LightCycler®480. GAPDH was used as an endogenous control, and only one peak of the melting curve of each reaction could be regarded as a valid result. The experiment was repeated three times. Primer sequences used in this study are shown in Supplementary File 1.
Hisscript 3 rt supermix for qpcr gdna wiper kit
Manufactured by Vazyme
Sourced in China
HisScript® III RT SuperMix for qPCR (+gDNA wiper) is a one-step reverse transcription and real-time PCR reagent kit. It contains a reverse transcriptase enzyme, a DNA polymerase, and a gDNA wiper component to remove genomic DNA contamination.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using hisscript 3 rt supermix for qpcr gdna wiper kit
1
RNA Extraction and qPCR Analysis
2
Comprehensive HCC Transcriptome Analysis
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In this study, total RNA was isolated by homogenizing the obtained clinical HCC samples using Trizol reagent (Invitrogen, USA). Then, the extracted RNA was reverse transcribed into cDNA by the HisScript® III RT SuperMix for qPCR (+gDNA wiper) kit (Vazyme, China). The qPCR experiments were performed according to the instructions of the ChamQ Universal SYBR qPCR Master Mix kit (Vazyme, China). MiRNA reverse transcription and qPCR experiments were performed using the Bulge-Loop miRNA qRT-PCR Starter kit (RiboBio, China). Roche Light-Cycler®480 was used to perform qPCR. GAPDH and U6 were used as endogenous controls for mRNA and miRNA, respectively. Primers used in this experiment were designed and purchased from GENEray (Shanghai, China) (Table S1 ).
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