Direct fluorescein labeling of DNA

U12201 at uicvm.uic.edu U12201 at uicvm.uic.edu
Wed Mar 22 14:52:00 EST 1995


Pierce Chemical has a system called OligoLink which converts
the 5' Phosphate to a primary amine. Once that is done, you
can react the NH2-DNA with NHS-Fluorescein (also Pierce) and
you have what you were looking for. The kit is sold in two
parts, one is the above described part, the other is activated
alk. phos. which will latch on the the activated DNA. The first
kit actually gives you a choice of making NH2 or SH respectively.
In article <edbeaty-2103951544380001 at koniskyj3.life.uiuc.edu>, edbeaty at uxa.cso.uiuc.edu (Edward Beaty) says:
>
>I'm wondering if it's possible to directly (non-enzymatically) link
>fluorescein to DNA.  I know you can incorporate fluoresceinated (?)
>nucleic acids into DNA by Klenow or terminal transferase, but I was
>wondering if there's a simpler chemical method using fluorescein
>isothiocyanate or another deriviative.  Also, if it is possible, has
>anyone tried to use this DNA for hybridizations i.e. Southern blots?
>Thanks for your help.
>
>Edward Beaty
>Department of Microbiology
>University of Illinois-Urbana
>
>--
>Ed Beaty
>Ed_Beaty at qms1.life.uiuc.edu
>"A mime is a wonderful thing to waste."



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