(4 pts) You are running an SDS-PAGE gel and want to load a total of 25 ug of pro
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Question
(4 pts) You are running an SDS-PAGE gel and want to load a total of 25 ug of protein in each well. Complete the table below indicating what volume of cach sample should be loaded. Well Sample 2 3 Weight Nuclcarexract Cytoplasmie Purified protein standard extract Total protein (ug) N/A 25 25 25 Sample Concentration (ug/ul) Loading volume (ul) N/A 50 5 ul (2 pts) Explain why it is important that you never touch an electrophoresis unit while it is running (2 pts) Oops! You forgot that your gel was running and haven't checked on it in a while. When you check the gel, you see that the sample is still sitting in the wells - it hasn't moved! The electrophoresis unit is plugged in and turned on. Give two reasons that this could happern.Explanation / Answer
Concentration = mass(ug)/volume(ug) ----> c =m/v
for 2 : c = 1 ug/ul, m = 25 ug
v = m/c = 25 ul
For 3 : c = 5 ug/ul, m = 25 ug
v = 25/5 = 5 ul
For 4 : c = 50 ug/ul, m = 25 ug
v = 25/50 = 0.5 ul
The electrophoresis unit should never be touched because it has Acrylamide, which is toxic to the human nervous system.
Reasons why the sample is still in the wells :
(1) Sample buffer such as glycerol hasn't been added to the sample, whose function is to increase the density of the sample so that it sinks to the bottom of the gel.
(2) The proteins weren't denatured properly into individual polypeptides, owing to which they can't pass through the pores in cross linked polymer of acrylamide because of their large size.
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