“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
Quantification of Mitochondrial DNA
“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
Corresponding Organization :
Other organizations : Weizmann Institute of Science
Variable analysis
- Mitochondrial DNA (mtDNA) quantification
- Relative mtDNA copy number
- Total RNA isolation using NucleoSpin RNA II kit
- Reverse transcription of 1 μg total RNA into cDNA using Bio-RT, dNTPs, and random hexamer primers
- QRT-PCR performed on Step One Plus, ABI instrument using SYBR Green PCR Master Mix
- Normalization of specific gene values to HPRT housekeeping gene using the ΔΔCt method
- Total DNA isolation using Quick-gDNA MiniPrep
- Quantification of mtDNA with SYBR green PCR Master Mix on Step One Plus, ABI instrument
- Calculation of relative mtDNA copy number from the subtraction of mtDNA copies to nuclear DNA copies
- Calculation of relative fold change based on the ΔΔCt method
- Not specified
- Not specified
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