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Southern blot with mtDNA?

Maria C.Costanzo mcc at proteome.com
Thu Apr 6 07:33:39 EST 2000


Dear Jukka,

	Southerns on total yeast miniprep DNA work very well with
mitochondrial probes because the mito genes are in high copy relative to
nuclear genes. Often you can see the mitochondrial bands on the ethidium
bromide-stained gel. I haven't worked in the lab for a few years so I can't
give you experimental details, but I used to do this all the time using the
standard Southern procedure and it worked very well; there's definitely no
need to purify mtDNA.

Regards,
Maria


>I'm preparing a Southern blot to see if there are any deletions in the
>yeast mtDNA and wondering what would be a good probe for this experiment. I
>quess nuclearDNA contamination is a big problem in mtDNA Southerns if you
>don't first extract the mtDNA with CeCl gradient to really purify it from
>total DNA? I just thought to make minilysates (as described in Yeast, a
>practical approach, Ed. Campbell and Duffus) and do the Southerns with
>these. Suggestions would be highly appreciated!
>
>Cheers,
>Jukka
>
>---


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Maria C. Costanzo, Ph.D.			Email mcc at proteome.com
Editor, Fungal Knowledge Systems 		Tel (607) 272-0687
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