Quantitating Radioactive label in DNA and RNA

John Stiles stiles at uhunix.uhcc.Hawaii.Edu
Wed Jun 29 13:32:03 EST 1994


In article <2us9ns$eso at matt.ksu.ksu.edu> hlhinkel at ksu.ksu.edu (Heidi Lanora Hinkel) writes:
>	I am looking for methods with which to quanitate the
>amount of P32 -ATP incorperated by end-labeling with T4 kinase,
>in both DNA and RNA probes. I would like to hear of any methods
>that you know of and which one you think is best. Thanks in
>advance
>		Heidi Hinkel
>		hlhinkel at matt.ksu.ksu.edu
>		Division of Biology
>		Kansas State University
>		Manhattan, KS 66506
>
Heidi,
   The easiest way I've found is to use DEAE membranes (Schleicher & Schuell
NA45 membranes).  Just spot some of the Rx on the filter and wash with 5%
Na2HPO4.  The nucleotides are washed off but RNA, DNA etc. is retained.  
This method gives low background and is quite fast.  
The original reference, that used Whatman DE81 chromotography paper,
is Litman, R.M. (1968) J. Biol. Chem. 243:6222-6233. 

john
stiles at uhunix.uhcc.hawaii.edu



More information about the Methods mailing list