Podcast
Questions and Answers
In the developing of SDS-PAGE a researcher has forgotten to add ______ to the loading buffer, it is likely that...
In the developing of SDS-PAGE a researcher has forgotten to add ______ to the loading buffer, it is likely that...
glycerol
A researcher wants to analyze a protein of 200kDa molecular mass. An appropriate separating gel would have a final acrylamide percentage of:
A researcher wants to analyze a protein of 200kDa molecular mass. An appropriate separating gel would have a final acrylamide percentage of:
- 1% (correct)
- 5%
- 20%
- 7%
The function of correcting the densitometry by using the value of the loading control is to:
The function of correcting the densitometry by using the value of the loading control is to:
- normalize the signal to obtain a ratio that allows to increase the specific signal
- normalize the signal, to obtain a ratio between the change caused by the experiment and a known and relatively stable signal value (correct)
- normalize the signal to obtain a ratio to decrease the background
- normalize the signal, to obtain a ratio between the densitometry value and the biological changes
An important variable when selecting restriction sites to initially open a plasmid would be:
An important variable when selecting restriction sites to initially open a plasmid would be:
A researcher is looking to generate a stable cell line, what type of vector would you advise using?
A researcher is looking to generate a stable cell line, what type of vector would you advise using?
One way to ensure the correct expression of a eukaryotic protein (without post-translational modifications) in a prokaryotic system could be to generate a cloning experiment from:
One way to ensure the correct expression of a eukaryotic protein (without post-translational modifications) in a prokaryotic system could be to generate a cloning experiment from:
A researcher wants to analyze a protein of 200kDa molecular mass. An appropriate concentrating gel would have a final acrylamide percentage of:
A researcher wants to analyze a protein of 200kDa molecular mass. An appropriate concentrating gel would have a final acrylamide percentage of:
A researcher seeks to clone the insulin gene to produce recombinant insulin and to purify it in order to use in his/her experiments of glucose regulation. The sequence of interest to insert into the appropriate vector should be:
A researcher seeks to clone the insulin gene to produce recombinant insulin and to purify it in order to use in his/her experiments of glucose regulation. The sequence of interest to insert into the appropriate vector should be:
A researcher is analyzing by Western Blot (WB) the phosphorylation of AKT. As for blocking he/she should:
A researcher is analyzing by Western Blot (WB) the phosphorylation of AKT. As for blocking he/she should:
A researcher is about to generate a figure from Western Blot (WB) results in a siRNA experiment. In one lane, there is a control siRNA (that do not knockdown any gene) and in the other lane, there is a siRNA positive treatment. However, researcher observe differences in the signal of the loading controls. Knowing that WB is semiquantitative, the approximation you could propose to compose the figure would be:
A researcher is about to generate a figure from Western Blot (WB) results in a siRNA experiment. In one lane, there is a control siRNA (that do not knockdown any gene) and in the other lane, there is a siRNA positive treatment. However, researcher observe differences in the signal of the loading controls. Knowing that WB is semiquantitative, the approximation you could propose to compose the figure would be:
In a WB experiment, blocking of the membrane was not optimal, thus is likely that:
In a WB experiment, blocking of the membrane was not optimal, thus is likely that:
In terms of ethics in scientific image manipulation, the next one is generally considered a good practice:
In terms of ethics in scientific image manipulation, the next one is generally considered a good practice:
The correct position of the layers of the WB "sandwich" starting from close to the negative electrode and moving towards the positive electrode is:
The correct position of the layers of the WB "sandwich" starting from close to the negative electrode and moving towards the positive electrode is:
One way to purify recombinant proteins obtained in prokaryotic systems could be to perform a cloning...
One way to purify recombinant proteins obtained in prokaryotic systems could be to perform a cloning...
Flashcards
What happens if glycerol is not added to the loading buffer in SDS-PAGE?
What happens if glycerol is not added to the loading buffer in SDS-PAGE?
Glycerol helps the sample sink to the bottom of the well, without it, the sample may float and disperse, leading to uneven band intensity and poor resolution during electrophoresis.
What acrylamide percentage is suitable for separating a 200kDa protein?
What acrylamide percentage is suitable for separating a 200kDa protein?
A higher percentage acrylamide gel is needed to separate larger proteins. A 20% gel would allow for better resolution of the 200kDa protein.
Why is densitometry normalized using a loading control?
Why is densitometry normalized using a loading control?
It adjusts the signal measured for the protein of interest to a known and stable value, like a loading control, to account for any potential variations in sample loading or protein expression in different experimental conditions.
Why is a unique restriction site important for plasmid cloning?
Why is a unique restriction site important for plasmid cloning?
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What kind of vector is suitable for creating a stable cell line?
What kind of vector is suitable for creating a stable cell line?
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How can you express a eukaryotic protein in a prokaryotic system?
How can you express a eukaryotic protein in a prokaryotic system?
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What acrylamide percentage is appropriate for a concentrating gel for a 200kDa protein?
What acrylamide percentage is appropriate for a concentrating gel for a 200kDa protein?
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What sequence should be used for cloning the insulin gene?
What sequence should be used for cloning the insulin gene?
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What is the purpose of blocking in Western Blot?
What is the purpose of blocking in Western Blot?
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How should a figure be composed for Western Blot experiment with siRNA treatment?
How should a figure be composed for Western Blot experiment with siRNA treatment?
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What happens if the blocking step in a Western Blot is not optimal?
What happens if the blocking step in a Western Blot is not optimal?
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What is considered a good practice in scientific image manipulation?
What is considered a good practice in scientific image manipulation?
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What is the correct position of the layers in a Western Blot sandwich?
What is the correct position of the layers in a Western Blot sandwich?
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How can you purify recombinant proteins obtained in prokaryotic systems?
How can you purify recombinant proteins obtained in prokaryotic systems?
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Study Notes
Biomedicine Methods and Techniques Exam
- SDS-PAGE Buffer: A researcher forgot glycerol in the loading buffer, likely leading to issues.
- Protein Analysis (200kDa): A 20% acrylamide separating gel is suitable for analyzing a 200kDa protein.
- Loading Control Normalization: Normalizing signal using a loading control creates a ratio to assess the experiment's effect against a standard signal, aiding data interpretation.
- Restriction Sites in Plasmids: Uniquely situated restriction sites prevent plasmid recircularization, crucial for DNA insertion without unnecessary loops.
- Stable Cell Lines: Using a plasmid vector with neomycin resistance is recommended for stable cell line generation.
- Eukaryotic Protein Expression: Cloning eukaryotic cDNA into a bacterial backbone can ensure proper protein expression without post-translational modifications.
- Concentrating Gel (200kDa): A 20% acrylamide gel is ideal for concentrating a 200kDa protein.
- Western Blot Blocking: Using skim milk (5%) is a common and usually effective way of blocking a Western Blot to avoid undesired protein interactions.
- Western Blot (WB) siRNA Experiments: To analyze siRNA experiments with a WB, correcting measured signals against loading controls yields comparable results.
- Western Blot Membrane Blocking: Insufficient membrane blocking results in improper signal-antibody interactions.
- Ethical Image Manipulation: Appropriately highlighting results without manipulating data in a way that misrepresents findings is key.
- Western Blot Sandwich Layers: The correct order during a Western blot is: sponge, filter paper, membrane, gel, filter paper, sponge.
- Recombinant Protein Purification: Using a specific tag and subsequent immunoprecipitation helps purify recombinant proteins.
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Description
Test your knowledge on essential biomedicine methods and techniques including protein analysis, SDS-PAGE, and stable cell line generation. This quiz covers crucial concepts such as loading controls, plasmid restriction sites, and eukaryotic protein expression. Prepare to deepen your understanding of experimental techniques in biomedicine.
