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Questions and Answers
Which sequence should be inserted into the appropriate vector for insulin gene study?
Which sequence should be inserted into the appropriate vector for insulin gene study?
In Western Blot analysis, which blocking agent has been suggested due to its variable effectiveness?
In Western Blot analysis, which blocking agent has been suggested due to its variable effectiveness?
When preparing a Western Blot figure from siRNA results, what is a potential issue the researcher faces?
When preparing a Western Blot figure from siRNA results, what is a potential issue the researcher faces?
Which of the following was not a proposed option for blocking in Western Blot?
Which of the following was not a proposed option for blocking in Western Blot?
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How does the choice of blocking agent affect Western Blot results?
How does the choice of blocking agent affect Western Blot results?
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What type of vector is specifically identified as a third generation with a particular tag?
What type of vector is specifically identified as a third generation with a particular tag?
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What is necessary to obtain a eukaryotic protein in a prokaryotic system?
What is necessary to obtain a eukaryotic protein in a prokaryotic system?
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For cloning the insulin gene, what is an important aspect for ensuring production of recombinant insulin?
For cloning the insulin gene, what is an important aspect for ensuring production of recombinant insulin?
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What would be the appropriate acrylamide percentage for an appropriate concentrating gel to analyze a 200kDa protein?
What would be the appropriate acrylamide percentage for an appropriate concentrating gel to analyze a 200kDa protein?
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Which option best describes the importance of using a bacterial backbone in cloning experiments?
Which option best describes the importance of using a bacterial backbone in cloning experiments?
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In which scenario would using eukaryotic mRNA be preferable for producing proteins?
In which scenario would using eukaryotic mRNA be preferable for producing proteins?
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What is a limitation of plasmid vectors when compared to viral vectors?
What is a limitation of plasmid vectors when compared to viral vectors?
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When cloning the insulin gene, which feature is critical for functionality in its expressed form?
When cloning the insulin gene, which feature is critical for functionality in its expressed form?
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What is an appropriate method to improve the accuracy of signal quantification in a WB experiment?
What is an appropriate method to improve the accuracy of signal quantification in a WB experiment?
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What consequence is most likely from suboptimal blocking of the membrane in a WB experiment?
What consequence is most likely from suboptimal blocking of the membrane in a WB experiment?
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Which of the following practices is considered ethical image manipulation in scientific research?
Which of the following practices is considered ethical image manipulation in scientific research?
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In the construction of a WB sandwich, which order correctly represents the positioning from the negative electrode to the positive electrode?
In the construction of a WB sandwich, which order correctly represents the positioning from the negative electrode to the positive electrode?
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What effect would an improperly blocked membrane have on the WB results?
What effect would an improperly blocked membrane have on the WB results?
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What is the likely consequence of forgetting to add glycerol to the loading buffer in SDS-PAGE?
What is the likely consequence of forgetting to add glycerol to the loading buffer in SDS-PAGE?
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Which of the following corrections is less advisable when dealing with WB signal quantification?
Which of the following corrections is less advisable when dealing with WB signal quantification?
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For analyzing a protein of 200kDa molecular mass, which final acrylamide percentage is most appropriate for the separating gel?
For analyzing a protein of 200kDa molecular mass, which final acrylamide percentage is most appropriate for the separating gel?
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Under which condition would performing partial manipulations of WB images be considered unethical?
Under which condition would performing partial manipulations of WB images be considered unethical?
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Why might discarding a WB experiment be considered a poor practice?
Why might discarding a WB experiment be considered a poor practice?
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What is the purpose of correcting densitometry using the loading control value?
What is the purpose of correcting densitometry using the loading control value?
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When selecting restriction sites to open a plasmid, what is an important variable to consider?
When selecting restriction sites to open a plasmid, what is an important variable to consider?
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What is a common consequence of having higher background in WB results?
What is a common consequence of having higher background in WB results?
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What type of vector is generally recommended for generating a stable cell line?
What type of vector is generally recommended for generating a stable cell line?
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In which scenario is it acceptable to manipulate imaging data in a WB study?
In which scenario is it acceptable to manipulate imaging data in a WB study?
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When designing an experiment, why is it important to include a loading control in Western blot analysis?
When designing an experiment, why is it important to include a loading control in Western blot analysis?
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What is an initial consideration when constructing a plasmid for cloning purposes?
What is an initial consideration when constructing a plasmid for cloning purposes?
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In electrophoresis, if a researcher uses an incorrect acrylamide concentration, what is a likely outcome?
In electrophoresis, if a researcher uses an incorrect acrylamide concentration, what is a likely outcome?
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What consequence does selecting a multi-restriction site have during molecular cloning?
What consequence does selecting a multi-restriction site have during molecular cloning?
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Why is it necessary to normalize densitometry values when analyzing protein expression?
Why is it necessary to normalize densitometry values when analyzing protein expression?
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Study Notes
Biomedicine Methods and Techniques Exam
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SDS-PAGE Buffer: If glycerol is omitted from the loading buffer, it's likely the sample will not load and run correctly.
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Protein Molecular Mass Analysis (SDS-PAGE): A 200 kDa protein requires a 20% separating gel for proper analysis.
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Densitometry Correction: Normalizing the signal with a loading control aids in analyzing results by providing a ratio for comparison between experimental and control signal intensities
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Plasmid Restriction Sites: Choosing unique restriction sites prevents recircularization, crucial for targeted insertion
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Stable Cell Line Vectors: Using plasmid vectors with antibiotic resistance (e.g., neomycin) is beneficial for creating stable cell lines.
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Eukaryotic Protein Expression in Prokaryotes: Using a eukaryotic cDNA with a bacterial backbone is a method used to produce eukaryotic proteins in prokaryotic systems.
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Concentrating Gel for Protein Analysis (SDS-PAGE): 200kDa protein analysis necessitates a 20% concentrating gel.
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Insulin Gene Cloning: To produce recombinant insulin, the mRNA sequence (as cDNA) is necessary for cloning into a vector.
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Western Blot Blocking Procedure: Skim milk (5%) is often used for blocking in Western Blotting experiments, as an alternative to BSA. It effectively prevents non-specific binding of antibodies to the membrane.
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Western Blot Loading Control Variations: If loading controls vary across lanes, a semi-quantitative densitometry approach is appropriate, using the control lanes for normalization and calculation.
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Western Blot Membrane Blocking: Inadequate membrane blocking can lead to high background in the experiment, due to the non-specific binding of the antibodies.
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Ethical Image Manipulation: Performing partial manipulations is generally considered an unethical means to highlight a result. Adjustments should be for data clarity and to prevent distortion.
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Western Blot Sandwich Assembly: The correct assembly order for a Western Blot sandwich, placing layers from negative to positive electrode, is sponge, filter paper, membrane, gel, filter paper, and sponge.
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Recombinant Protein Purification: Cloning with a specific tag, followed by immunoprecipitation, is a common method for purifying recombinant proteins from prokaryotic systems.
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Description
Test your knowledge on essential biomedicine methods and techniques. This quiz covers topics such as SDS-PAGE, protein analysis, plasmid vectors, and eukaryotic protein expression. Evaluate your understanding of various laboratory practices in biomedicine.
