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ivanzaharov [21]
3 years ago
15

The breaking buffer that we use this week contains 10mM Tris, pH 8.0, 150mM NaCl. The elution buffer is breaking buffer that als

o contains 300mM imidazole. Describe how the instructor made the 0.25L elution buffer for all the students this week given 500ml of 1M of Tris (121.1 g/mole) (pH8.0), 750ml of 5M NaCl (MW = 58.44 g/mole) and 350ml of 1M imidazole (68.08 g/mole). Show your calculations for credit.
Chemistry
1 answer:
olasank [31]3 years ago
5 0

Answer:

A breakdown of the breaking buffer was first listed with its respective component and their corresponding value; then a table was made for the stock concentrations in which the volume that is being added was determined by using the formula M_1*V_1 = M_2*V_2. It was the addition of these volumes altogether that make up the 0.25 L (i.e 250 mL)  with water

Explanation:

Given data includes:

Tris= 10mM

pH = 8.0

NaCl = 150 mM

Imidazole = 300 mM

In order to make 0.25 L solution buffer ; i.e (250 mL); we have the following component.

Stock Concentration             Volume to be             Final Concentration

                                               added            

1 M Tris                                     2.5 mL                         10 mM

5 M NaCl                                  7.5 mL                        150 mM

1 M Imidazole                           75 mL                         300 mM    

M_1*V_1 = M_2*V_2. is the formula that is used to determine the corresponding volume that is added for each stock concentration

The stock concentration of Tris ( 1 M ) is as follows:

M_1*V_1 = M_2*V_2.

1*V_1= 0.01 M *250mL\\V_1 = 2.5mL

The stock concentration of NaCl (5 M ) is as follows:

M_1*V_1 = M_2*V_2.

1*V_1= 0.15 M *250mL\\V_1 = 7.5mL

The stock concentration of Imidazole (1 M ) is as follows:

M_1*V_1 = M_2*V_2.

1*V_1= 0.03 M *250mL\\V_1 = 75mL

Hence, it is the addition of all the volumes altogether that make up 0.25L (i.e 250 mL) with water.

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