Efficient Buffer Preparation: 500ml MgCl2, Glucose, ATP, and Tris-Cl at pH 7.8

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SUMMARY

The discussion focuses on preparing a 500ml reaction buffer containing 0.82% MgCl2, 62.5 mM glucose, 950 nM ATP, and 100 mM Tris-Cl at pH 7.8. Calculations for the required stock solutions were provided, including 28 mL of 15% MgCl2, 17.7 mL of 1.75M glucose, and 0.08 mL of 5mM ATP. The participants debated whether to use an existing pH 8.5 Tris solution or to prepare a new buffer using the Henderson-Hasselbalch equation. The consensus leans towards preparing a new buffer for accurate pH adjustment.

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Homework Statement


Prepare 500ml of a reaction buffer containing .82% MgCl2, 62.5 mM glucose, 950 ATP, and 100mM Tris -Cl, pH 7.8 Use the stock solutions you have prepared

we had to prepare in previous problems:

500ml 1.75M glucose
25mL 2.5% glucose
2L of 1.8 Tris, pH 8.5


Homework Equations


C1V1 = C2V2
Pka Tris = 8.1
glucose mol mass = 180.16
ATP mol mass = 507.8

------------


The Attempt at a Solution



my calculation for MgCl2:

.82% = .82g/100ml = 8.2g/L
15% = 15g/100ml = 150g/L

C1V1=C2V2

150*x = 8.2*500

x = 28mL stck 15% MgCl2

My calculation for glucose

1.75M glucose = 315g/L
62.5mM = 11.2 g/L

C1V1=C2V2

11.2*500 = 315*x

x = 17.7 ml 1.75M glucose stock

Here is my calculation for ATP

5mM ATP = 3g/L
950nM ATP = .00048g/L

C1V1=C2V2

3*x = .00048*500

x =.08ml of stocK 5mM ATP

My calculation for the Tris I am not sure about, can I use my original pH 8.5 solution the prof asked me to prepare, or do I have to start over with the Tris acid and base components and use henderson H equation? does this all look right so far?
 
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C1V1 = C2V2 works for molar concentrations as well, you don't have to convert them to g/L.

My guess is you should prepare new buffer from HH equation.
 
Borek said:
C1V1 = C2V2 works for molar concentrations as well, you don't have to convert them to g/L.

My guess is you should prepare new buffer from HH equation.

is the rest of it right?
 
Looks OK, but I just skimmed.
 

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