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Table 1 Calculation of heteroplasmy of LCLs

From: Patient-derived lymphoblastoid cell lines harboring mitochondrial DNA mutations as tool for small molecule drug discovery

Panel (A)
GM00333 (wildtype) Ct(wildtype) − Ct(mutant) % mutant mtDNA
3460G>A (ND1) − 9.92 ± 0.08 0.1 ± 0.01
11778G>A (ND4) − 10.47 ± 0.15 0.07 ± 0.01
8993T>G (ATP6) − 9.93 ± 0.48 0.13 ± 0.04
  1. (A) ARMS-qPCR was used to quantify wildtype or mutant forms of mtDNA in the 3 loci shown using total DNA isolated from GM00333 (wildtype) LCLs. Heteroplasmy was calculated based on Ct values. GM00333 (wildtype) cells have < 0.13% mutant mtDNA in either of the loci analyzed
  2. (B) Similarly, heteroplasmy was calculated for LCLs known to harbor mtDNA mutations at the loci indicated. As a control, DNA isolated from GM11605 or GM00333 were mixed in the ratios shown prior to qPCR, and yields expected heteroplasmy values
  3. Average ± SEM for three independent experiments are shown
Panel (B)
Strain Ct(wildtype) − Ct(mutant) % mutant mtDNA
GM11605 (ND1_3460G>A) 12.42 ± 0.45 99.98 ± 0.01
GM10742 (ND4_11778G>A) 8.75 ± 0.57 99.71 ± 0.06
GM10744 (ND4_11778G>A) 8.99 ± 0.05 99.8 ± 0.01
GM13741 (ATP6_8993T>G) 6.94 ± 0.61 98.81 ± 0.39
GM13740 (ATP6_8993T>G) 7.76 ± 0.55 99.37 ± 0.2
GM11605:GM00333 (1:1) 0.7 ± 0.27 61.64 ± 4.46
GM11605:GM00333 (1:5) − 2.05 ± 0.2 19.61 ± 2.16
  1. (A) ARMS-qPCR was used to quantify wildtype or mutant forms of mtDNA in the 3 loci shown using total DNA isolated from GM00333 (wildtype) LCLs. Heteroplasmy was calculated based on Ct values. GM00333 (wildtype) cells have < 0.13% mutant mtDNA in either of the loci analyzed
  2. (B) Similarly, heteroplasmy was calculated for LCLs known to harbor mtDNA mutations at the loci indicated. As a control, DNA isolated from GM11605 or GM00333 were mixed in the ratios shown prior to qPCR, and yields expected heteroplasmy values
  3. Average ± SEM for three independent experiments are shown