Blood samples (8.5 ml) are collected in PAXgene Blood DNA Tubes, where they may be stored or transported. For DNA isolation, the blood is transferred to processing tubes (supplied already filled with cell lysis buffer), and the solution is mixed to lyse red and white blood cells. Cell nuclei and mitochondria are pelleted by centrifugation, washed, and resuspended in digestion buffer. Protein contaminants are removed by incubation with a protease. DNA is precipitated in isopropanol, washed in 70% ethanol, dried, and resuspended in resuspension buffer.
Multiplex PCR of fragments from the mitochondrial genes tRNAlys/ATPase (0.92 kb), tRNAleu(UUR) (0.63 kb), and ND4 (0.29 kb), using 250 ng DNA from 8 donors as starting material.
Genomic DNA isolated from 8 blood donors using the PAXgene Blood DNA System. [A] Agarose gel analysis; [B] pulsed-field gel electrophoresis for enhanced separation of high-molecular-weight genomic DNA. M: markers.