You will need to make 50X TAE so that you can do gel electrophoresis. One of the

Question

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?

Determine the amounts/volumes of the chemicals needed to prepare the following solutions:  

5. 100 ml of 0.1 N HCl from concentrated (12.0 M) HCl [For HCl, 1 N = 1 M]

6. 100 ml of 1.0 M Tris pH = 7.6.   MW of Tris base is 121 g/mole.

7. pH 7.6 is close to a neutral pH. When making 1.0 M Tris base, would you expect to need HCl or NaOH to decrease the pH to 7.6?

Explanation / Answer

Components required

EDTA -50 mM-372.24 g/mol(MW)

Tris-1M- 121.14 g/mol(MW)

glacial acetic acid 1M- 60

Preparation of a 0.5 M EDTA stock solution

2

For 100 ml:

preparation of 50x buffer

weight 12.11g tris base and dilute in 80 ml Deionized water then make up to 100ml

Q.7 there is no need to adjust ph

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