This is probably a very basic question, but I've been thinking about this lately. I've routinely done RNase A treatments without a buffer for 3h. The constructs I'm looking to isolate very stable so I don't generally worry about the pH. I noticed that a lot of protocols ask for a buffer to be used. I've noticed pH of these buffers is anywhere between 5-8. Because of this range, I'm wondering what the role of the buffer is. Is it as simple as maintaining the pH at a range where the ribonuclease is best functional?
Dear Lisa
Each enzyme has a pH range beyond witch the specific activities are sub optimal. In some case you will have to add metal ions for maximal activity.
Indeed the reason to use a buffer is to ensure optimal pH range for the enzyme activity.
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