The detailed method was described previously49 (link). All primers and probes were ordered
from IDT (Coralville, Iowa). The 3D digital PCR was used according to the
manufacturer’s protocol. QuantStudio™ 3D Digital
PCR Master Mix v2 and individual QuantStudio 3D digital PCR 20K Chip kit v2
were purchased from Thermo scientific (Applied Biosystems, Waltham, MA).
Prepared sample mix was loaded into ChIP using QuantStudio 3D Digital PCR
Chip Loader (Thermo scientific). Chip PCR amplification was performed in a
ProFlex PCR System (96 °C for 10 min; 39 cycles of 60 °C for 2
min and 98 °C for 30 sec; and 60 °C for 2 min). After
amplification, each chip was loaded into the QuantStudio 3D Digital reader.
Data were analyzed using QuantStudio 3D Analysis Suite (Thermo scientific).
All biological repeats were performed in at least triplicate.
from IDT (Coralville, Iowa). The 3D digital PCR was used according to the
manufacturer’s protocol. QuantStudio™ 3D Digital
PCR Master Mix v2 and individual QuantStudio 3D digital PCR 20K Chip kit v2
were purchased from Thermo scientific (Applied Biosystems, Waltham, MA).
Prepared sample mix was loaded into ChIP using QuantStudio 3D Digital PCR
Chip Loader (Thermo scientific). Chip PCR amplification was performed in a
ProFlex PCR System (96 °C for 10 min; 39 cycles of 60 °C for 2
min and 98 °C for 30 sec; and 60 °C for 2 min). After
amplification, each chip was loaded into the QuantStudio 3D Digital reader.
Data were analyzed using QuantStudio 3D Analysis Suite (Thermo scientific).
All biological repeats were performed in at least triplicate.
ΔmtDNA primers F: TTGCTTTTTCTTTATATGTTTTG; R:
TTTATTTAATTTGGTTAAACAAGAGGT. ΔmtDNA probes 5’
6-FAM/ZEN/3’ IBFQ:
/56-FAM/AGGATCGTA/ZEN/ACATTTTATTTTTTTGCTTTA/3IABkFQ/.
Wildtype primers F: GCTTTTTCTTTATATGTTTTGTG.
R: TCACCTTCAGAAAAATCAAATGG wildtype mtDNA probes
5’ HEX/ZEN/3’ IBFQ:
/5HEX/AATTATAGT/ZEN/AATTGCTGAACTTAACCGGGC/3IABkFQ/
