Total RNAs were isolated by TRIzol reagent (Takara, Dalian, China) according to the manufacturer’s instruction. gDNA was isolated by Genomic DNA Isolation Kit (Sangon Biotech, Shanghai, China). The quality and quantity of RNA and DNA were detected by Nanodrop 2000 spectrophotometer (Thermo Fisher Scientific, USA). The nuclear and cytoplasmic fractions were purified by PARIS Kit (Ambion, Life Technologies). RNA was reverse transcribed by HiScript II Q RT SuperMixfor qPCR (+gDNA wiper) (Vazyme, Nanjing, China). The AmpliTaq DNA Polymerase (Life Technologies) was used for PCR. The 2% agarose gel electrophoresis was performed to observe the cDNA and gDNA PCR products. AceQ qPCR SYBR Green Master Mix (Vazyme, Nanjing, China) was used for qRT-PCR, and GAPDH was used to normalize the level of circRNA and mRNA. Hydrolysis probe-based RT-qPCR assay of miRNA was performed according to the manufacturer’s instructions (Applied Biosystems). The miRNA level was normalized by small nuclear U6. Primers are listed in Supplementary Table S3
Hydrolysis probe based rt qpcr assay of mirna
Manufactured by Thermo Fisher Scientific
The Hydrolysis probe-based RT-qPCR assay of miRNA is a laboratory equipment product designed for the detection and quantification of microRNA (miRNA) molecules. This assay utilizes reverse transcription followed by quantitative polymerase chain reaction (RT-qPCR) technology, with hydrolysis probes as the detection method.
Automatically generated - may contain errors
Lab products found in correlation
Aceq qpcr sybr green master mix, by Vazyme (2 mentions)
Hiscript 2 q rt supermix for qpcr gdna wiper, by Vazyme (2 mentions)
Paris kit, by Thermo Fisher Scientific (2 mentions)
Trizol reagent, by Takara Bio (2 mentions)
Amplitaq dna polymerase, by Thermo Fisher Scientific (2 mentions)
Nanodrop 2000 spectrophotometer, by Thermo Fisher Scientific (2 mentions)
Genomic dna isolation kit, by Sangon (2 mentions)
2 protocols using hydrolysis probe based rt qpcr assay of mirna
1
Comprehensive RNA and DNA Isolation Protocol
2
Comprehensive RNA and DNA Isolation Protocol
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Total RNAs were isolated by TRIzol reagent (Takara, Dalian, China) according to the manufacturer's instruction. gDNA was isolated by Genomic DNA Isolation Kit (Sangon Biotech, Shanghai, China). The quality and quantity of RNA and DNA were detected by Nanodrop 2000 spectrophotometer (Thermo Fisher Scientific, USA). The nuclear and cytoplasmic fractions were purified by PARIS Kit (Ambion, Life Technologies). RNA was reverse transcribed by HiScript II Q RT SuperMixfor qPCR (+gDNA wiper) (Vazyme, Nanjing, China). The AmpliTaq DNA Polymerase (Life Technologies) was used for PCR. The 2% agarose gel electrophoresis was performed to observe the cDNA and gDNA PCR products. AceQ qPCR SYBR Green Master Mix (Vazyme, Nanjing, China) was used for qRT-PCR, and GAPDH was used to normalize the level of circRNA and mRNA. Hydrolysis probebased RT-qPCR assay of miRNA was performed according to the manufacturer's instructions (Applied Biosystems). The miRNA level was normalized by small nuclear U6. Primers are listed in Table s3.
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