Flourescein labeling- Help!

Michele Fuortes mfuortes at med.cornell.edu
Wed Jun 16 18:18:00 EST 1993

Sheila Gay Buzzee writes:

>  Our lab uses a large amount of fluorescein-labeled primers, and we are 
>attempting to synthesize them ourselves instead of remaining at the mercy of 
>suppliers for pure, high-quality primers.  We have no problem making the 
>primers themselves, but we are having problems with labeling.
>  Does anyone know how to label the 3' or 5' ends of double-stranded DNA?  (Or 
>both?)  Does anyone have experience with flourescein-labeling primers?  200-bp 
>ds DNA?
>  Please send any primary references if possible, or advice if you don't have 

We have been using very succesfully the kit from Amersham to do 3'
labelling. The kit basically includes Terminal Transferase, fluorescinate
ATP and buffers, blocking agents etc. It's a "little" expensive ;-) but it
should not bet too complicated to just get TT and they also sell all the
different fluorescinated NTPs.

Hope this helps.


Michele Fuortes, MD
Dept. Cell Biology and Anatomy
Cornell University Medical College
New York, NY
E-Mail: mfuortes at med.cornell.edu

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