2.9. RNA isolation and quantitative real‐time PCR analysis
This protocol is extracted from research article:
Kir6.1 improves cardiac dysfunction in diabetic cardiomyopathy via the AKT‐FoxO1 signalling pathway
J Cell Mol Med, Feb 6, 2021; DOI: 10.1111/jcmm.16346

RNA from heart tissue or NRVMs was extracted with TRIzol reagent (Invitrogen, Carlsbad, CA, USA). cDNA synthesis was performed with a PrimeScriptTM RT regent Kit (Takara, Kyoto, Japan). Quantitative real‐time PCR (qRT‐PCR) was performed in duplicate in a total reaction volume of 25 μL using SYBR‐Green master mix (Takara) and conventional protocols. The primer sequences for qRT‐PCR are listed in Table S3. Expression was normalized to that of the housekeeping gene, 36β4. Quantitative data was calculated using the comparative CT method.

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