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DNA was extracted from peripheral blood using the QIAamp DNA Mini Kit (QIAGEN, Valencia, CA, USA), as previously described.24 All DNA samples were analyzed using whole exome sequencing25 and subsequently verified by Sanger sequencing of the mutations located in the OPA1 and SSBP1 genes. LHON diagnoses were confirmed by polymerase chain reaction using a MyCycler Thermal Cycler (Bio-Rad Laboratories, Hercules, CA, USA) and purification, followed by sequencing to confirm one of the three hotspots (m.11778G>A, m.14484T>C, or m.3460G>A).
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