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1) Prepare 1 L of 1X transfer buffer using the 10X stock

10X Transfer buffer (500 mL)

15.15 g Tris base + 72 g glycine

Fill to 500 mL with water

2) Calculate the molarity of Tris in the solution and the molarity of glycine in the 10X. The pH of the solution is 8.5. Then, calculate the concentrations of each component in the 1X solution.

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Related questions

TPE isa commonly used buffer for DNA electrophoresis. This buffercontains 89 mM Tris, 23 mM Phosphate, 2 mM EDTA as the final concentrations. The final pHis 8.3. The 10X stock solution (10 X the final concentration) isnormally prepared. You have the Tris powder (mol. wt. 121.14),Phosphoric acid H3PO4 (liquid 91.3 % pure,density 1.61 g/mL, mol. wt. 98) and a 0.5 M EDTA stock solution.You need to make up 1 L of 10X TPE. How would you make itup?

BUFFERS:

(Final concentration for 1x is 20 mM Tris; 500 mM NaCl; 1 mM CaCl2; 1 mM MgCl2; pH 7.4)

5x ConA Buffer, (1L)

• 700 ml dH2O, set stirring and add:

• 100 ml 1M Tris, pH 7.2-7.4

• 5 ml 1M MgCl2

• 5 ml 1M CaCl2

• 146 g NaCl

• Check pH is between 7.2 and 7.4

• To 1000 ml with dH2O

Elution Buffer A: ~0.2M AMM: 0.1 g alpha methyl mannopyranoside in 2 mls ConA buffer

Elution Buffer B: ~0.5M AMM: 0.2 g alpha methyl mannopyranoside in 2 mls ConA buffer

Elution Buffer C: ~1 M AMM: Add 0.4 g alpha methyl mannopyranoside in 2 mls ConA buffer (you may need more of this)

Formula weight for alpha methyl mannopyranoside is 194.2 g.

Questions:

Buffers: Calculate the concentrations of Tris, CaCl2 and MgCl2 in the 5x ConA Buffer stock

Tris:

CaCl2:

MgCl2:

For Buffer B: What is the exact concentration of the AMM to three significant figures

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 [you will be asked to make this solution as practice]

6. 100 ml of 1.0 M Tris pH = 7.6 MW of Tris base is 121 g/mole [you will be asked to make this solution as practice]

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 adjust the pH to 7.0?

To do electrophoresis, you will need to make a solution called TAE, which is a specific mixture of Tris base, acetic acid, and EDTA. Assuming no errors and no spills, you will need a minimum of 50 ml 1X TAE to make your agarose gel plus 1 liter of 1X TAE for gel running buffer. TAE is normally made as a 50X concentrated stock.

8. What is the minimum volume of 50X TAE that you will need to make?

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