Effects of ROS Damage on MtDNA Mutations in Oocytes during Female Reproductive Aging
To determine whether an increase in mitochondrial matrix ROS could cause a surge in mtDNA mutations in oocytes during female reproductive aging, mtDNA sequencing was conducted on oocytes with Sod2 depletion and mtSSB heterozygous knockout from females aged at three weeks, six months, and nine months, respectively. Using the rolling circle amplification technique, we successfully amplified adequate mtDNA content from individual oocytes of various females, allowing us to prepare a PCR-free library (to minimize PCR bias) for subsequent next-generation sequencing.