Mitochondria are double-membrane cytoplasmic organelles that generate over 90% of a cell’s chemical energy through respiration. The mitochondrial genome (mtDNA) is a circular, intron-free, double-stranded, haploid DNA molecule approximately 16.5 kb in length that encodes 37 genes. Unlike nuclear DNA, mtDNA exhibits a high rate of mutation, damage, and copy number variation, which has been closely linked to a variety of diseases, including diabetes, Alzheimer’s disease, and muscle disorders. To investigate these associations, it is essential to obtain high-quality mtDNA.

The RayBio^® Mitochondrial DNA Isolation Kit (Bead-Based) offers a simple and effective tool for isolating mtDNA (1.5 hours for 12 samples) from a variety of cells and tissues. This kit yields highly pure mtDNA, free of nuclear genomic DNA contamination, and is suitable for a wide range of downstream applications, including enzymatic manipulation, Southern blotting, cloning, PCR analysis, and amplification.

• Isolates high-quality mtDNA
• No nuclear DNA contamination
• High yield and purity
• Compatible with multiple cell types
• Easy to use

• Sterile nuclease-free pipette tips (barrier tips recommended) and microfuge tubes
• PBS
• Microcentrifuge
• Magnetic separator for microcentrifuge tubes (2 mL)
• Vortex mxer
• Water bath or dry heat block capable of heating to 56°C
• 100% ethanol (30 mL)
• Precision pipettes to deliver 2 µL to 1 mL volumes
• Tissue homogenizer if using tissue samples (Dounce tissue grinder recommended)
• Refrigerated centrifuge with swing out rotor and refrigerated microcentrifuge

Contact our technical support team for questions about compatibility: techsupport@raybiotech.com

1. Prepare cell or tissue samples as instructed in the manual.
2. Lyse cells with Cell Lysis Buffer 1 for 15 min and Cell Lysis Buffer 2 for another 2 minutes on ice.
3. Lyse mitochondria with Protease K and mtDNA Lysis Buffer at 56°C for 10 minutes.
4. Bind mtDNA with magnetic beads at RT for 10 minutes.
5. Wash with Wash Buffers A, B, and C.
6. Elute mtDNA with Elution Buffer.
7. mtDNA can be used immediately or stored at or below -20°C.

Store Cell Lysis Buffer and Protease K at 4°C. Store the rest of the components in a dry environment between 2 - 30°C. The kit can be stored for up to 12 months.