I am working with 30bp DNA and want to study the Hyperchromicity of it. So, basic idea is to study the absorbance of DNA under UV. As i am studying it wrt temperature (22°-80°C) should i expose the sample (DNA+TBE) continuously to the UV (but it can possibly damage the DNA). Is there any limit of UV exposure to DNA so that the DNA is not damaged.
Hello Ms. Kaur,
TO study the effect of UV dose you will have to know its degradation components which can be analyzed after delivering the UV dose. For the basic idea, you can deliver different UV doses to your sample for ex. 50,100, 200 mj/cm2. and study the changes in absorbance at that particular wavelength. greater the change, greater the DNA damage. If you do not want to denature the DNA, test the doses that do not bring about any changes in absorbance. The accuracy of dose measurement is of key importance. Good luck.
You can go through my paper for reference if needed. Patulin degradation and cytotoxicity evaluation of UV irradiated apple juice using human peripheral blood mononuclear cells: CHANDRA et al.
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