DNA laddering - apoptosis - any suggestion about the extraction procedure ? (Oct/09/2008 )
HI!
I've just posted a topic on the cell biology forum...I'm a very beginner on DNA extraction.
In few words, I'm currently using the Wizard kit from Promega (find the attached protocol) to extract DNA which requires
RNase treatment
protein precipitation
DNA isolation
Should I use a different kit made expressely for fragmented DNA? If DNA is correctely fragmented, would it be easy to see ladder? Or are there some other tricks I should know to see any result?? Up to now, in the same experiment I have
1.weak smear
2.smear
3. big gDNA band
4.nothing
5. big gDNA band
Where 1 is control, 3 and 5 positive controls for apoptosis, 2 and 4 positive controls for apoptosis at lower apoptotic agent concentration...
Thank you!! I'm surrounded by MDs, nobody knows anything about molecular biology!!
Did you ever try Trizol? Or home made Phenol-Chlorophorm extraction?