Tritiated Thymidine and The Rate of DNA Syn.

Ronald P. Herzig, Jr. r-herzig at nwu.edu
Fri Oct 6 21:04:29 EST 1995


--This is a very simple inquiry for all of you tritium experts out there.  I am growing 
cells in culture and adding 1 micro Ci of 3H per mL of media for four hours.  Upon 
harvesting the cells and isolating the DNA, I am interested in quantitating the amount of 
3H incorporated relative to other harvestings.  In this manner I wish to assess the rate of 
DNA synthesis.  I am looking for technical suggestions on quantitating 3H levels.  Many of 
you will respond by simply telling me to use scintillation.  Fine, but I need to know more 
than that.  I need to know the type of vial and cocktail used if it is different than that 
used for any other isotope, and I need to know how much DNA you add to the vial and whether 
or not it is on a glass fiber filter or on nylon, etc.  In otherwords please just don't 
tell me to use scintillation.  I figured that much out for myself.  I need 
specific technical advise.  And for those who know anything about detection with 
autoradiography, please contribute.  Thanks in advance.

Ronald P. Herzig, Jr.
Northwestern University, Evanston, IL.   USA
r-herzig at nwu.edu




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