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After restriction digestion my bands just disappear


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#1 mab

mab

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Posted 02 December 2004 - 05:14 AM

I need some urgent help here guys....


I did mini-prep of my plasmid clones using alakali-lysis method...yield close to 400- 700 ng/ul... good bands on the gel

Now to identify poitive clones I did restriction digestion with pvuII for one set of clones and HindIII for the next set. As control, I also digested a vector.

and I ran the digested plasmid side-by-side to the undigested ones....


and the digested ones have just disappeared: all i see is a smear-type band at the bottom of the gel. The undigested ones are fine and the control was also cut.

Is it that I have some nucleases in my miniprep or what?

Has anyone else faced this problem...I'd appreciate some help here

thanks

mab




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