Nucleic Acid Electrophoresis Process
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Nucleic Acid Electrophoresis Process

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Questions and Answers

What is the primary advantage of capillary electrophoresis over gel electrophoresis?

  • Increased sensitivity and immediate detection
  • Decreased runtime and preparation time
  • Automatic analysis of results by software
  • All of the above (correct)
  • What is the purpose of a buffer system in electrophoresis?

  • To carry the current and protect the samples
  • To maintain the pH of the solution
  • To prevent severe fluctuations in the pH of the gel
  • All of the above (correct)
  • Which of the following is a characteristic of a buffered solution?

  • The buffer concentration must be low to provide sufficient acidic and basic forms
  • The addition of acid or base will significantly affect the pH
  • The buffer does not protect sample molecules from damage
  • The pH remains constant as the buffer molecules take up or release protons (correct)
  • What is the main disadvantage of capillary electrophoresis?

    <p>Requirement for fluorescent labeling of samples</p> Signup and view all the answers

    How does a buffer maintain the pH of a solution near its pKa?

    <p>By providing sufficient acidic and basic forms to buffer the solution</p> Signup and view all the answers

    What is the role of buffer ions in the electrophoresis process?

    <p>To carry the current through the gel and prevent pH fluctuations</p> Signup and view all the answers

    What is the purpose of inserting a comb into the gel in gel electrophoresis?

    <p>To create holes for loading the sample</p> Signup and view all the answers

    Why is it crucial to keep the thickness of the gel and the volume of the buffer constant in gel electrophoresis?

    <p>To ensure consistent results by maintaining current flow</p> Signup and view all the answers

    What will happen if contact is made with the gel buffer while the current is on in gel electrophoresis?

    <p>There will be a significant shock hazard</p> Signup and view all the answers

    How does the charge/mass ratio of sample molecules affect their migration in gel electrophoresis?

    <p>It determines the speed at which samples migrate through the gel matrix</p> Signup and view all the answers

    What happens to the gel after the comb is removed and it is placed into the gel box in gel electrophoresis?

    <p>The gel is allowed to cool and solidify</p> Signup and view all the answers

    Why are gel baths designed to stop the current when a protective cover is removed from the gel bath?

    <p>To prevent shock hazards when handling the gel bath</p> Signup and view all the answers

    What is the role of methanol and acetic acid after electrophoresis on the sample?

    <p>Fix the sample</p> Signup and view all the answers

    How does silver staining work under optimal pH conditions?

    <p>Deposits metallic silver on the target</p> Signup and view all the answers

    What is the main difference between silver diamine and silver nitrate procedures?

    <p>Silver nitrate precipitates black silver salt upon introduction of formaldehyde</p> Signup and view all the answers

    Which dye sits on the minor groove of the double helix for double-stranded DNA staining?

    <p>SYBR green I</p> Signup and view all the answers

    What is a disadvantage of SYBR gold dye?

    <p>Stains both DNA and RNA</p> Signup and view all the answers

    How does SYBR green differ from EtBr in terms of binding to DNA bases?

    <p>SYBR green intercalates between bases</p> Signup and view all the answers

    What is the impact of raising the buffer concentration on gel stability?

    <p>Lessens gel stability</p> Signup and view all the answers

    Which ions move more quickly through the gel?

    <p>+2, -2, +3</p> Signup and view all the answers

    Why are Tris base and Borate preferred for maintaining constant pH during electrophoresis?

    <p>They remain partly uncharged at desired pH</p> Signup and view all the answers

    Which buffer is prone to precipitation and could result in localized distortions in DNA electrophoresis?

    <p>TBE</p> Signup and view all the answers

    What other characteristics, besides pKa and charge, should be considered when choosing a buffer?

    <p>Toxicity, interaction with other components</p> Signup and view all the answers

    How does an increase in buffer concentration impact the conductivity of the electrophoresis system?

    <p>Increases conductivity</p> Signup and view all the answers

    What is the main reason that the nucleic acid should be completely denatured (single stranded) before separation?

    <p>To allow the nucleic acid to migrate according to its size rather than its secondary structure</p> Signup and view all the answers

    What is the primary purpose of using fluorescent labels on the nucleic acids?

    <p>To allow the nucleic acids to be detected during the capillary electrophoresis process</p> Signup and view all the answers

    What is the main advantage of using a capillary in the electrophoresis process?

    <p>Improved heat dissipation properties</p> Signup and view all the answers

    Which of the following is NOT a common forensic application of the capillary electrophoresis technique described in the text?

    <p>Protein structure analysis</p> Signup and view all the answers

    How does the electrokinetic injection process work to draw the sample into the capillary?

    <p>A transient high-positive charge is applied to the platinum electrode close to the end of the capillary</p> Signup and view all the answers

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