PREPARING LABORATORY

SOLUTIONS AND REAGENTS II

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Solutions With
More Than One Solute
RECIPE I

Na2HPO4
6g
KH2PO4
3g
0.4% glycerol 10 mL
Dissolve in water.
Bring to a volume of 1 liter.
Recipe lists amounts of solutes.
This is easy to follow

Solutions With
More Than One Solute Cont
RECIPE II

1 M MgCl2
0.1 M Tris
Gives the final concentration of each solute.
You must calculate the amount of each to use.
This is harder to interpret

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A Recipe With
Multiple Solutes, SM Buffer
SM BUFFER
0.1 M NaCl
1 mM MgSO4
0.2 M Tris, pH 7.5
0.01% gelatin

This might be whole recipe as given in a manual

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Strategy 1:
PREPARING SM BUFFER WITHOUT STOCK
SOLUTIONS, OVERVIEW:

Prepare a solution of 0.2 M Tris, pH 7.5.

Calculate grams of each of the other solutes
required.
Weigh out solutes and dissolve directly in Tris
buffer.

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Strategy 1

Decide how much buffer to make, for

example, 1 liter.
1 liter of 0.2 M Tris requires 24.2 g of Tris
base. (MW = 121.1 g/mole.)
Dissolve Tris in about 700 mL of water and
bring pH to 7.5.
Do not bring Tris to volume.

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4. 1 liter of 0.1 M NaCl requires 5.84 g of NaCl. Add

this to Tris buffer.
5. 1 liter of 1 mM MgSO4 requires 1/1000 of its MW.
MgSO4 comes in more than one hydrated
form; read
container to determine MW.
Weigh out correct amount.
Add it to Tris buffer.

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0.01% gelatin is 0.1 g in 1 L. Weigh out and

add to Tris buffer.
Stir to dissolve
BTV 1 L
Check and record pH.

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Strategy 2:
PREPARING SM BUFFER WITH STOCK
SOLUTIONS

In overview:
Four solutes are each prepared separately as
concentrated stock solutions.

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When the four stocks are combined, they

dilute one another to the proper final
concentrations

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Steps Are:
1.

Prepare a stock solution of Tris buffer at pH

7.5.
a. No set rule as to what should be
concentration of stock.
b. Example: To make 1 L of 1 M stock,
dissolve 121.1 g of Tris base in about 900
mL of water.

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2.

c.
Bring pH to 7.5.
d.
BTV 1L.
Prepare a stock solution of magnesium
sulfate, for example, 1 M.
a. To make 100 mL, dissolve 0.1 MW of
MgSO4 in water, bring to a volume of 100 mL.

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3.

Prepare a stock solution of NaCl, for

example,1 M.
a. To make 100 mL of stock, dissolve 5.84
grams in water and BTV 100 mL.

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4.

Prepare a stock solution of gelatin, for

example, 1%.
a. Dissolve 1 g in a final volume of 100 mL
of water.

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5.

To make the final solution, combine the right

amounts of each stock.
a. Since this is a situation where stocks are
diluted, use the C1V1 = C2V2 equation
four times, once for each solute.

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For example, to calculate how much

Tris stock is required:
C1 V1 = C2
V2
(1 M) (?)
(0.2 M) (1000 mL)
? = 200 mL

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Summary
To make 1000 mL of SM buffer, combine:
200 mL
1 mL
100 mL
10 mL

Tris stock
MgSO4 stock
NaCl stock
Gelatin stock

BTV 1L; check and record pH.

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Which Strategy?

Both strategy 1 and 2 are correct.

Generally efficient to make stock solutions of
frequently used solutes because weighing out
chemicals is time-consuming.

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