Explain how you would prepare 10 mL of the Laemli Sample Loading Buffer as given below,...
Explain how you would prepare 10 mL of the Laemli Sample Loading Buffer as given below, using the following stock solutions: 1 M DTT, 10% SDS, 1M Tris HCl pH 6.9, 100% Glycerol, 0.2% stock of Bromophenol Blue.
Homework Answers
Answer
Preparation of 10 mL of the 1 X Laemli Sample Loading Buffer
(a) SDS
Stock Concentration (M1) : 10 %
Working Concentration (M2) : 4%
Final volume (V2) : 10ml
Initial Volume: V1
According to molarity formulae
M1V1 = M2V2
10 X V1 = 4 X 10
V1 = 40 /10 = 4 ml
(b) Tris HCl pH 6.9
Stock Concentration (M1) : 1 M
Working Concentration (M2) : 0.125M
Final volume (V2) : 10ml
Initial Volume: V1
According to molarity formulae
M1V1 = M2V2
1 X V1 = 0.125 X 10
V1 = 1.25 ml
(c) Glycerol
Stock Concentration (M1) : 100 %
Working Concentration (M2) : 20 %
Final volume (V2) : 10ml
Initial Volume: V1
According to molarity formulae
M1V1 = M2V2
100 X V1 =20 X 10
V1 = 2ml
d) Bromophenol Blue
Stock Concentration (M1) : 0.2 %
Working Concentration (M2) : 0.004%
Final volume (V2) : 10ml
Initial Volume: V1
According to molarity formulae
M1V1 = M2V2
0.2 X V1 = 0.004 X 10
V1 = 0.2ml
e) DTT
Stock Concentration (M1) : 1 M
Working Concentration (M2) : 50mM
Final volume (V2) : 10ml
Initial Volume: V1
According to molarity formulae
M1V1 = M2V2
1 X V1 = 50 X 10
V1 = 500 / 1000 ( 1M = 1000mM)
V1 = 0.5ml
So,
- SDS : 4ml
- DTT : 0.5ml
- Bromophenol Blue: 0.2ml
- Glycerol : 2ml
- Tris HCl pH 6.9 : 1.25ml
Total volume : 10 - 7.95= 2.05 ml (Water)
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Explain how you would prepare 10 mL of the Laemli Sample
Loading Buffer as given below,...
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