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how to move for ligation with low conentration of digested plasmid - (Apr/27/2006 )

I am trying to clone in pGBKT7 vector but the vector concentration is too low. I proceeded for a double digestion using EcoRI and BamHI sites but now i am confused wheather I shall gel elute or ethanol precipitate or follow any other method to remove restriction enzymes from the mixture and proceed for ligation with minimum loss of DNA. Please suggest me.

-Rash-

The problem is if you perform ethanol precipitation you also precipitate the enzymes... so you should heat inactivate them or you could clean up your digestion with a pcr clean up kit.
By the way, what is the concentration of your vector?

-Fedex-

hi
i agree with fedex. A kit is more reliable to purify. Then elute in a great volume (100µl and pellet with adding 1ml buthanol and vortex well. wash twice with 1ml etoh70% and resuspend in 10µl

-fred_33-