To prepare a tris buffer for the DNase activity testing and to determine the concentration of DNA in the DNA stock to be used for activity testing.
Determining the Concentration of the DNA stock
- We want to add 200 ng DNA to each sample for activity testing.
- the absorbance and concentration of DNA are linearly related according to the equation:
unknown [DNA]/A260 = 50 ug/ml/1.0
- Took a UV-vis measurement of the DNA by putting 2 uL of DNA and 198 uL water in a small volume cuvette
- Plotted the data and found the absorbance at 260 nm
- Solved the above equation and found the concentration of DNA to be 1610 ug/mL
- Then figured out the volume of DNA stock that contains 200 ng → there are 200 ng in 0.124224 uL. Therefore, we will add 1 uL of DNA to each sample
Tris Buffer Preparation
- A 50 mM tris buffer with a pH of 7.6 was prepared by
- Add data and results here...
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