no band after digestion - (Feb/09/2008 )

i pcr my dna , run gel, gel extraction, digest , run gel
everything is ok except the gel running at last
i can only my vector, my dna is gone

the buffer and the enzyme should be fine, cause the vector band is very clear
and i have checked the step of gel extraction, after extraction i run the gel again with the product from gel extraction, which is fine

so now my problem is i can not find my dna after i digested it

waiting for help....thanks

I think because your dna was too little.

i have added much more dna but still cannot find it.....

maybe the digestion is not inactivated properly?

hi, not sure i understand , are you digesting your PCR product?
i dont know what gel extraction method you are using, but with QIAgen you can use for purification without having tu run a gel. follow the same protocol but mix buffer QG with your digestion mix.

I am confused. IF its the PCR product which you are missing, how do you see the vector? Vector must not be there.

I agree that sometime columns are not very good and one could lose most of DAN to a column. Try repeating it and hopefully you have a good amount of PCR product for the experiments.