Measurements of the amount of biotin conjugated to DNA
harold.taylor.delete-this at remove-this.uni-tuebingen.de
Fri Feb 9 08:52:36 EST 2001
sleboda at completegen.com ("Jowita Sleboda") wrote:
... snip ...
>I have a lambda DNA which I have tagged with biotin. Now I want to measure the
>amount of biotin that has bound to the lambda DNA. I thought about using streptavidin
>conjugated with alkaline phosphatase and p-nitrophenyl phosphate and measure it
>spectrophotometrically, but then I have problems with the DNA which has to be
>immobilized first. Any suggestions?
if you have the resources use radioactively labeled avidin /
streptavidin. Immobilize your DNA on a membrane, block it and toss the
radioactivity onto it. Wash, count and voila! Enzymatic tests require
a certain calibration. You can do it but it's more work.
My 2 cents,
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