Total RNA was isolated from tissue and cells with use of TRIzol reagent (Invitrogen), and 0.5 μg RNA was converted to cDNA with the SuperScript II Reverse Transcriptase kit (Invitrogen). Primers for CYP2C8, 2C9, 2J2, sEH and β-actin were as we described previously [17 (link)]. qRT-PCR amplification involved the PrimeScript Real-Time RT-PCR reagent kit (Takara Biotechnology [DALIAN] Co.) and Applied Biosystems Prism 7300 (Invitrogen). DNase-treated RNA was amplified without reverse transcriptase as a negative control. Human hepatocellular carcinoma tissue RNA was a positive control and water was a blank control. Amplification of β-actin was an internal control. The relative expressions of CYPs and sEH were normalized to their corresponding normal control tissue.
Primescript real time rt pcr reagent kit
Manufactured by Takara Bio
The PrimeScript Real-Time RT-PCR reagent kit is a set of reagents used for performing reverse transcription and real-time PCR amplification of RNA samples. The kit includes all the necessary components to convert RNA into cDNA and amplify the resulting DNA in a real-time PCR reaction.
Automatically generated - may contain errors
2 protocols using primescript real time rt pcr reagent kit
1
Quantitative RT-PCR Analysis of CYP and sEH Genes
2
Quantitative RNA Expression Analysis
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Total RNA was isolated from cultured cells by using Trizol reagent (Invitrogen) following the manufacturer's protocol. cDNA was synthesized from 1.0 μg total RNA with the PrimeScript Real-Time RT-PCR reagent kit (Takara Biotechnology [DALIAN] Co.) as previous described [25 (link),26 (link)]. The samples were amplified by using the 7300 Realtime PCR System (Applied Biosystems). Relative mRNA expression was analyzed according to the comparative Ct method and normalized to that of β-actin. The primer sequences are listed in Table S1.
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