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

Explanation:

From the given information ;the objective is to determine how the instructor made the 0.25L elution buffer

0.25 L elution buffer = 250 mL elution butter

The breaking buffer that we use this week contains

10mM Tris    =   0.01 M

150mM NaCl  =   0.15 M

300mM imidazole.  = 0.3 M

The stock concentration  of Tris in 1M

Therefore ; by using the formula: ; we can determine the volume in the preparation; so;

In NaCl, The amount of stock concentration is 5 M

so; using the same formula; we have:

From Imidazole ; the amount of stock concentration is

Thus; we can have a table as shown as :

Stock concentration        volume to be added        Final concentration

1 M of Tris                              2.5 mL                            10 mM

5 M of  NaCl                          7.5 mL                             150 mM

1 M of Imidazole                    75  mL                            300  mM

In conclusion. the addition of all the volume make up the 250 mL elution buffer that is equivalent to 0.25 L.

the product of the pressure and volume is also constant