Mitochondrial DNA sequencing of homo sapiens: OSCC H103 cell line (Amplicon)
Mitochondrial DNA (mtDNA) was extracted using a QIAprep Miniprep Kit (QIAGEN, Germany). The mtDNA fraction was enriched and size-selected using solid-phase reversible immobilization (SPRI) paramagnetic beads (Agencourt AMPure XP; Beckman Coulter Inc., USA). Two long PCR products with tail sequences, which spanned the entire mtDNA, were amplified using AtMax Taq DNA Polymerase (Vivantis Technologies, Malaysia) on an Arktik Thermal Cycler (Thermo Scientific Inc., USA). The DNA libraries were prepared using the 1D Ligation Sequencing Kit (SQK-LSK108; Oxford Nanopore Technologies, UK). During the library preparation, an additional limited PCR was performed using barcoded primers complementary to the tail sequences previously added in the long PCRs (PCR Barcoding Kit, EXP-PBC001; Oxford Nanopore Technologies, UK).
H103_Amplicon