Quantification of DNA in presence of NADPH

nauduri dhananjaya nauduri at hotmail.com
Mon Mar 26 21:50:07 EST 2001

I working on microsomal activation of benzene. For the in vitro reaction, I 
use Microsomes, NADPH (co-factor), Calf thymus DNA and benzene. After 
completion of reaction NADPH is also co-precipitating along with DNA. Since, 
both DNA and NADPH show absorption at 260 nm, quantification of DNA is not 
possible to measure DNA concentration at 260 nm. (A conventional method of 
separating DNA from the reaction mixture by isoamyl alcohol+chloroform and 
precipitation by cold ethanol would give both DNA+NADPH.)

Diphenylamine method is not also possible because perchloric acid break both 
DNA and NADPH to produce deoxyribose moiety.

I request anybody suggest me a method(s). I appreciate any easy way to 
eliminate NADPH without disturbing the DNA or simple spectrophotometric 
estimation would be great. I have no access to fluorescence 
spectrophotometer.  I sincerely appreciate for your advise. Email me 
:nauduri at hotmail.com
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