Hi!

I've been trying to isolate complete mtDNA plasmids from cheek cells (16.5kbp). Once the nuclear DNA is pelleted and the supernatant containing the mtDNA is transfered in a new tube, pelleted, cleaned and resuspended, which would be the best way to go to get ONLY mtDNA in my sample, and no remaining nuclear DNA? Doing a sucrose gradient, or using a spin column for plasmids? Or something else altogether?

Please, do not tell me to follow the directions on the kit: I am not using one ;)

Any complete protocol for mtDNA purification (no kits) that has been tested by one of you would be truly welcome, and any opinion on which technique works best too :)

Little update: I do not have access to an ultracentrifuge (16 500xg max), nor to liquid nitrogen.

Thank you very much, and for those in the US, have a good Memorial Day week end!