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Phenol Chlorform Extractions - (Oct/20/2005 )

I am trying to use an alkaline lysis procedure with a phenol chloroform extraction.

I keep ending up with low molecular weight bands on my gel (about 500bp) and no evidence of any high molecular weight DNA. I was told to make sure my phenol was fresh so I bought a brand new bottle of equilibrated phenol pH 8.0 and tried again with the same small band results. Does anyone have any suggestions of why I can't get any DNA from my samples? My samples are fresh nematodes. Never been frozen, never been near ETOH.

thanks for any suggestions!



I`m doing this type of extraction too, but not with alkali lysis...
Perhaps you pipette too axious... That has been my mistake at the beginning...
Do you clean up your phe:chl-DNA mixture with chl? You should...



your band appears as kind of a smear?
If so that's RNA...