Question

"You suspect that a chemical carries out oxidative DNA Cleavage. Outline chemical carries out oxidative DNA cleavage. Outline the experiments that you would use to test this hypothesis in vitro and how would you differentiate b/w oxidative and hydrolytic DNA cleavage

Answer 1

To verify oxidative DNA cleavage induced by a chemical, one could treat isolated DNA with the suspect compound and search for oxidative base damage, differentiating this from hydrolytic cleavage with control treatments.

Step-by-step explanation:

To test the hypothesis that a chemical induces oxidative DNA cleavage, in vitro experiments could involve treating isolated DNA with the chemical and analyzing the resulting DNA fragments. Detecting oxidative cleavage specifically can be achieved by looking for oxidized bases such as 8-oxo-7,8-dihydroguanine (8-oxoG). These oxidation products can be identified by chromatography or mass spectrometry. To differentiate between oxidative and hydrolytic DNA cleavage, one could compare the treated DNA with DNA subjected to alkaline or acidic conditions known to cause hydrolytic cleavage.

If the cleavage pattern is different in the presence of the chemical, suggesting the formation of oxidized bases rather than simple cuts in the DNA backbone, this would indicate oxidative cleavage. Additionally, the use of scavengers or inhibitors that specifically prevent oxidative damage could serve as control treatments to support the results.