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Hi everybody,
I am trying to subclone my gene from 3kb vector to 6 kb vector.my insert size is 210bp... for insert release i digesting 60000 ng of my 3kb vector to get minimum of 300ng insert.this i have to use for ligation after agarose elution.problem while digestion in my reaction i somehow my DNA concentration getting reduced...i could see only 1/10th of concentration after digestion.i noticed, in my digestion reaction mixture i am getting white precipitate....reaction components dna 6000ng, tris 10mm, mgcl2 10mm, bsa100ng/ml(added sparetely). if anybody have idea or suggestion it will be helpful...