Question

How are we getting the volumes in this solution for example for 10Xs buffer how did we get 1 microliter please explain for each reagent how the microliters were obtained; ddH20, 10X buffer A, 1 microgram/microliter DNA stock, and enzyme at 10 U/microliter.

Question: What is the most concentrated stock of buffer A you can make? Now if you want to prepare an enzymatic reaction containing 1X buffer A, 100 ng/ul DNA stock and 1 Unit/ul (U/ul) of enzyme proceed as follows: First: decide how much reaction volume you want. Second: calculate what volume of each reagent is required. So if you decide a 10 ul reaction volume is sufficient then proceed as follows (assuming DNA starting stock is at 1 μg/μ| and enzyme is at 10 Uhl): pipetted with a P10) (pipetted with a P2) (pipetted with a P2) (pipetted with a P2) 7 pul 1 ul ddH20 10X buffer A 1 μg/μ| DNA stock Enzyme at 10 U/pl (Total Reaction volume) 1 ul 1011

Answer 1

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