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GLP Practical 1a Calculation 2022 BCHN213 - MEMO

The digestion buffer consists of: - 1.03 g NaCl - 2.25 mL 250 mM Tris-Cl solution - 10.84 g EDTA - 69.23 mL 13% sodium dodecyl sulfate solution - 8.75 mL proteinase K solution These components would be weighed/measured out and dissolved in approximately 600-700 mL water. The pH would be adjusted if needed and more water added to reach the final volume of 750 mL.

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KAGISO BRIAN MOTSHUPHI
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62 views3 pages

GLP Practical 1a Calculation 2022 BCHN213 - MEMO

The digestion buffer consists of: - 1.03 g NaCl - 2.25 mL 250 mM Tris-Cl solution - 10.84 g EDTA - 69.23 mL 13% sodium dodecyl sulfate solution - 8.75 mL proteinase K solution These components would be weighed/measured out and dissolved in approximately 600-700 mL water. The pH would be adjusted if needed and more water added to reach the final volume of 750 mL.

Uploaded by

KAGISO BRIAN MOTSHUPHI
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© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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You are supplied with the following:

NaCl (Mr = 58.443 g/mol)


250 mM Tris-Cl, pH 8 solution (1
Litre) EDTA, sodium salt (Mr = 380.2
g/mol) 13 % Sodium dodecyl sulfate
solution
Proteinase K solution (150 mg dissolved in 1 ml ddH2O)
You need a digestion buffer consisting of the following:
23.5 mM NaCl
750 uM Tris-Cl, pH 8
38 mM EDTA, pH 8
1.2 % Sodium dodecyl sulfate
1.75 mg/ml proteinase K
Calculate:
How will you prepare 750 ml of the digestion buffer? Show all your steps and calculations
and remember to explain how exactly you will make it up.
Answer Point Value:18 points
Model Short Answer:--------
Feedback:
NaCl: 23.5 mM = 0.0235 M; Mr = 58.443 g/mol
C=n / V
n=Cx
V (1)
n = 0.0235 M x 0.75 L
n = 0.017625
mol (1)
n= m / Mr
m=nx
Mr (1)
m = 0.017625 mol x 58.443 g/mol
m = 1.03
g (1)
Tris-Cl, pH 8: 750 uM = 0.00075 M
C1V1 = C2V2 (1)
(0.25 M)(V1) = (0.00075 M)(750 mL)
V1 = 0.5625 / 0.25
V1 = 2.25
mL (1)
EDTA, pH 8: 38 mM = 0.038 M; Mr = 380.2 g/mol
n=Cx
V (1)
n = 0.038 M x 0.75 L
n = 0.0285
mol (1)
n= m / Mr
m=nx
Mr (1)
m = 0.0285 mol x 380.2 g/mol
m = 10.84
g (1)
Sodium dodecyl sulphate solution:
C1V1 = C2V2 (1)
(13%)(V1) = (1.2%)(750 mL)
V1 = 900 / 13
V1 = 69.23
mL (1)
Proteinase K solution:
C1V1 = C2V2 (1)
(150 mg/mL)(V1) = (1.75 mg/ml)(750 mL)
V1 = 1312.5 / 150
V1 = 8.75 mL (1)

Remember to explain exactly how you will make it up


The salts need to be weighed off first and dissolved in some water (1). The liquids then
need to be measured and added to the solution (1). Water then needs to be added to the
solution (approx. 600ml/700ml), adjust pH if needed (1) and add more water to give a final
volume of 750 mL. (1)

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