Total RNA from cells was isolated using the NucleoSpin RNA II kit (Macherey-Nagel, Duren, Germany). A concentration of 1 μg aliquot of the total RNA was reverse transcribed into cDNA using Bio-RT (Bio-Lab, Jerusalem, Israel), dNTPs and random hexamer primers. qRT-PCR was performed on Step One Plus, ABI instrument (Applied Biosystems, Grand Island, NY, USA) using SYBR Green PCR Master Mix (Quanta BioSciences, Gaithersburg, MD, USA). The values for the specific genes were normalized to HPRT housekeeping gene using the ΔΔCt method.
“qPCR analysis for mitochondrial DNA” has been published previously [13 (link)][14 (link)],[15 (link)],. Total DNA was isolated using Quick-gDNA MiniPrep (Zymo Research, Cat. No. D3007). Mitochondrial DNA (mtDNA) was quantified with SYBR Green PCR Master Mix on Step One Plus, ABI instrument (Applied Biosystems, Grand Island, NY, USA). The relative mtDNA copy number was calculated from the subtraction of mtDNA copies to nuclear DNA copies. The relative fold change was then calculated based on the ΔΔCt method. Primers used in this study are shown in TableS3 .
“qPCR analysis for mitochondrial DNA” has been published previously [13 (link)][14 (link)],[15 (link)],. Total DNA was isolated using Quick-gDNA MiniPrep (Zymo Research, Cat. No. D3007). Mitochondrial DNA (mtDNA) was quantified with SYBR Green PCR Master Mix on Step One Plus, ABI instrument (Applied Biosystems, Grand Island, NY, USA). The relative mtDNA copy number was calculated from the subtraction of mtDNA copies to nuclear DNA copies. The relative fold change was then calculated based on the ΔΔCt method. Primers used in this study are shown in Table
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