Biomedicine Methods and Techniques Exam
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Questions and Answers

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:

  • 1% (correct)
  • 5%
  • 20%
  • 7%
  • 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:

    <p>find a restriction site that is unique to prevent the plasmid from recircularization</p> Signup and view all the answers

    A researcher is looking to generate a stable cell line, what type of vector would you advise using?

    <p>plasmid vector with neomycin resistance</p> Signup and view all the answers

    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:

    <p>a eukaryotic cDNA and using a bacterial backbone</p> Signup and view all the answers

    A researcher wants to analyze a protein of 200kDa molecular mass. An appropriate concentrating gel would have a final acrylamide percentage of:

    <p>7%</p> Signup and view all the answers

    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:

    <p>the sequence of the mRNA (as cDNA)</p> Signup and view all the answers

    A researcher is analyzing by Western Blot (WB) the phosphorylation of AKT. As for blocking he/she should:

    <p>use BSA as its blocking action is usually better than skim milk</p> Signup and view all the answers

    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:

    <p>to densiometrate the bands and correct its values with the loading controls' signal</p> Signup and view all the answers

    In a WB experiment, blocking of the membrane was not optimal, thus is likely that:

    <p>the primary antibody will not interact properly</p> Signup and view all the answers

    In terms of ethics in scientific image manipulation, the next one is generally considered a good practice:

    <p>cutting out a portion of the total image, if the procedure performed is indicated</p> Signup and view all the answers

    The correct position of the layers of the WB "sandwich" starting from close to the negative electrode and moving towards the positive electrode is:

    <p>sponge, filter paper, membrane, gel, filter paper, sponge</p> Signup and view all the answers

    One way to purify recombinant proteins obtained in prokaryotic systems could be to perform a cloning...

    <p>with a specific tag and then performing an immunoprecipitation</p> Signup and view all the answers

    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.

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