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_________________________________________________________________
Get your FREE download of MSN Explorer at http://explorer.msn.com
---
