Mitochondrial DNA Copy Number Quantification

Total genomic DNA was extracted from LCLs and relative mitochondrial DNA (mtDNA) copy number was estimated by SYBR Green assay in a q-PCR system (Thermofisher). Cytochrome B (Cyt B) and NADH dehydrogenase 1 (ND1) genes were used to represent the mtDNA, and pyruvate kinase (PK) gene was used to represent the nuclear DNA. The primers selected for this experiment were based on an earlier study for mtDNA copy number by Gu et al.^{24 (link)}. Relative mtDNA copy numbers of the genes Cyt B and ND1, normalized to the single-copy nuclear gene PK and relative to the calibrator is given by 2^−ΔΔCt.

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Paul P., Iyer S., Nadella R.K., Nayak R., Chellappa A.S., Ambardar S., Sud R., Sukumaran S.K., Purushottam M., Jain S, & Viswanath B. (2020). Lithium response in bipolar disorder correlates with improved cell viability of patient derived cell lines. Scientific Reports, 10, 7428.

Publication 2020

q-PCR system

Assay Cytochrome b Gene Genomic Lcls Mtdna Nadh dehydrogenase 1 Primers Pyruvate kinase Sybr green

Corresponding Organization :

Other organizations : National Institute of Mental Health and Neurosciences, Institute for Stem Cell Biology and Regenerative Medicine, National Centre for Biological Sciences

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Variable analysis

independent variables

Genome extraction method (not explicitly mentioned)

dependent variables

Relative mitochondrial DNA (mtDNA) copy number
Expression of Cytochrome B (Cyt B) gene
Expression of NADH dehydrogenase 1 (ND1) gene

control variables

Pyruvate kinase (PK) gene (used as a single-copy nuclear gene reference)

controls

Positive control: Not specified
Negative control: Not specified

Annotations

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