Key features and details
- Assay type: Direct
- Assay time: 3 hr
- Sample type: Adherent cells, Suspension cells, Tissue
Product nameMitochondrial DNA Isolation Kit
Sample typeTissue, Adherent cells, Suspension cells
Assay time3h 00m
Mitochondrial DNA Isolation Kit ab65321 provides convenient tools for isolating mtDNA from a variety of cells and tissues in high yield and purity, without contaminations from genomic DNA. The purified mtDNA can be used for a variety of studies such as enzyme manipulations, Southern blotting, cloning, PCR analysis, and amplifications
Please note: One kit is sufficient for 50 samples if Adherent cell or Suspension cells are used and for 25 samples if tissue is used.
Mitochondrial DNA isolation protocol summary:
- collect cells by centrifugation at 600 x g for 5 min and wash with PBS
- centrifuge for 5 min and resuspend pellet in cytosol extraction buffer
- incubate on ice for 10 min
- homogenize in dounce tissue grinder
- centrifuge for 10 min at 700 x g to pellet nuclei and intact cells
- transfer supernatant and centrifuge at 10,000 g for 30 min
- remove supernatant, resuspend pellet in cytosol extraction buffer and centrifuge at 10,000 g for 30 min
- resuspend pellet (mitochondria) and lyse in mitochondrial lysis buffer for 10 min
- add enzyme mix and incubate for 60 min
- add ethanol and incubate for 10 min
- centrifuge for 5 min and retain pellet (mitochondrial DNA)
Mitochondria are semi-autonomous organelles which functions in aging process, apoptosis, anti-HIV drugs, and cancers. Mitochondrial DNA (mtDNA) has a very high mutation rate and the mutations on mtDNA appear to be related to certain diseases such as diabetes, Alzheimer's disease, and muscle disorders. Isolation and quantification of mtDNA are often required to study the relationships between the diseases and mtDNA.
Storage instructionsStore at -20°C. Please refer to protocols.
Components Identifier 1 kit 5X Cytosol Extraction Buffer 1 x 20ml Enzyme Mix (lyophilized) Red 1 vial Mitochondrial Lysis Buffer Purple 1 x 1.8ml TE Buffer 1 x 1.5ml
RelevanceMitochondria are semiautonomous organelles which function in the aging process, apoptosis, anti-HIV drugs, and cancers. Mitochondrial DNA (mtDNA) has a very high mutation rate and the mutations on mtDNA appear to be related to certain diseases such as diabetes, Alzheimer’s disease, and muscle disorders.
ab65321 has been referenced in 17 publications.
- Casciano JC et al. MYC regulates fatty acid metabolism through a multigenic program in claudin-low triple negative breast cancer. Br J Cancer 122:868-884 (2020). PubMed: 31942031
- Cai Y et al. Elimination of senescent cells by ß-galactosidase-targeted prodrug attenuates inflammation and restores physical function in aged mice. Cell Res N/A:N/A (2020). PubMed: 32341413
- Stampar SN et al. Linear Mitochondrial Genome in Anthozoa (Cnidaria): A Case Study in Ceriantharia. Sci Rep 9:6094 (2019). PubMed: 30988357
- Kaiser S et al. Neuroprotection after Hemorrhagic Stroke Depends on Cerebral Heme Oxygenase-1. Antioxidants (Basel) 8:N/A (2019). PubMed: 31635102
- Wang Y et al. Metformin Improves Mitochondrial Respiratory Activity through Activation of AMPK. Cell Rep 29:1511-1523.e5 (2019). PubMed: 31693892