My textbook states: “In order to cut the DNA with restriction enzymes, it needs to be in pure form, free from other macromolecules. Therefore DNA isolated from other chemicals of the protoplasm like carbohydrates, lipids and proteins via techniques of DNA isolation.” My question is, why does DNA need to be alone? Won’t restriction enzymes work if there were a DNA molecule in a sugar solution? I understand that the DNA has to be freed from cell membranes and walls for restriction enzymes reach the DNA but how does the presence of other macromolecules hinder DNA’s digestion of a restriction enzyme?
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$\begingroup$ AFAIK its not that other macromolecules hinder the activity of restriction enzymes, its just that we don't want other macromolecules in the solution, we want pure DNA for our job. I might be wrong though... $\endgroup$– another 'Homo sapien'Commented Oct 5, 2017 at 18:01
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Some other macromolecules or other solutes may be present that could interfere with the affinity of the restriction enzyme. For example, there could be salts that denature or modify the conformation of the enzyme or inhibitors present that may interfere with the enzyme. Hope this helps.
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$\begingroup$ Welcome to Biology.SE! Could you please add some references? $\endgroup$ Commented Oct 6, 2017 at 5:43
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$\begingroup$ No Problem! promega.com/-/media/files/resources/protocols/technical-manuals/… $\endgroup$– JacobCommented Oct 6, 2017 at 15:53