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13 Dec 2019

1) Calculate how many grams agarose should be added to 150 mL 1X TAE to make it 1% (mass/volume) agarose solution

2)Calculate final concentration and components in 1X TAE. The 50X TAE is 2.0 M Tris-Acetate, pH 7.6, 0.05 M EDTA

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Keith Leannon
Keith LeannonLv2
17 Dec 2019

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Related questions

You will need to make 50X TAE so that you can do gel electrophoresis. One of the ingredients is 0.5 M EDTA pH = 8.0. 
EDTA stands for disodium ethylenediamindetratraacetate•2(H2O). This EDTA solution will be provided to you. However, you should know how it was made, and the following two questions address this.

3. Determine the amount of EDTA needed to prepare 100 ml of 0.5 M EDTA pH = 8.0. The EDTA MW = 372.2 g/mole

4. As EDTA dissolves in water, the [H+] will increase (note that full name of EDTA includes "acetate"). To increase to a pH = 8.0, would you need to add HCl or NaOH?

1. a) Calculate how to prepare 10.0 mL of 12% separating gel. The separating gel is 12% acrylamide, 0.1% SDS, 0.375 M Tris-Cl, pH 8.8.

b) Calculate how to prepare 5.0 mL of 4% stacking gel. The stacking gel is 4% acrylamide, 0.1% SDS, 0.125 M Tris-Cl, pH 6.8.

c) How much 4X loading buffer needs to be added to each sample to have 1X loading buffer in each sample?

d) If each sample were to be the same volume of 40.0 µL, how much 4X loading buffer needs to be added to each sample to have 1X loading buffer in each sample?

e) Why are small proteins the last samples collected for gel filtration but the first to reach the bottom for gel electrophoresis?

4.) a.) Given a 25X stock solution of Tris Glycine SDS (i.e. running buffer in gel electrophoresis), what volumes of the stock and DI water should be combined to make 100 mL of a 1X Tris Glycine SDS solution?

b.) You have 1 mL of a previously prepared sample buffer to which you wish to add DTT (which is unstable in solution). What volume of 1M DTT will you need to add to achieve a final concentration of 50 mM DTT in the sample buffer?

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