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Which of the following was/were in the Primer Mix of our PCR activity? Select all that apply, and choosing incorrect answers will cause a penalty.

Question 21 options:

template DNA from an insect not infected with Wolbachia

primers for the insect gene

dNTPS

DNA polymerase

primers for the Wolbachia gene

template DNA from an insect infected with Wolbachia

Question 22 (3 points)

Which of the following was/were in the positive control tube of our PCR activity that was loaded in the thermocycler?

Select all that apply, and choosing incorrect answers will cause a penalty.

Select any reagent that was in the bead AND any reagent that was added to the tube.

Question 22 options:

template DNA from an insect infected with Wolbachia

template DNA from an insect not infected with Wolbachia

dNTPS

primers for the insect gene

DNA polymerase

primers for the Wolbachia gene

Question 23 (3 points)

Which of the following was/were in the negative control tube of our PCR activity that was loaded in the thermocycler?

Select all that apply, and choosing incorrect answers will cause a penalty.

Select any reagent that was in the bead AND any reagent that was added to the tube.

Question 23 options:

DNA polymerase

primers for the insect gene

template DNA from an insect infected with Wolbachia

primers for the Wolbachia gene

dNTPS

template DNA from an insect not infected with Wolbachia

Question 24 (3 points)

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Which of the following was/were in the NTC tube of our PCR activity that was loaded in the thermocycler?

Select all that apply, and choosing incorrect answers will cause a penalty.

Select any reagent that was in the bead AND any reagent that was added to the tube.

Question 24 options:

template DNA from an insect infected with Wolbachia

template DNA from an insect not infected with Wolbachia

primers for the Wolbachia gene

dNTPS

DNA polymerase

primers for the insect gene

1 answer

To answer the PCR-related questions accurately, let's break down each one based on common PCR practices:

Question 21: Primer Mix of PCR Activity

  1. template DNA from an insect not infected with Wolbachia - Typically not included unless specifically studying non-infected samples.
  2. primers for the insect gene - Usually included if the insect gene is being amplified.
  3. dNTPS - Required for the PCR reaction, so this would be included.
  4. DNA polymerase - Essential for PCR, so this would be included.
  5. primers for the Wolbachia gene - Included if Wolbachia is the target for amplification.
  6. template DNA from an insect infected with Wolbachia - Included if the goal is to amplify genes from Wolbachia-infected DNA.

Correct Answers:

  • Primers for the insect gene
  • dNTPS
  • DNA polymerase
  • Primers for the Wolbachia gene
  • Template DNA from an insect infected with Wolbachia

Question 22: Positive Control Tube of PCR Activity

In a positive control tube, they typically include:

  1. template DNA from an insect infected with Wolbachia - Yes, this ensures that the PCR process can properly amplify the intended target.
  2. template DNA from an insect not infected with Wolbachia - Typically not included for a positive control unless it’s meant to validate the insect gene amplification without Wolbachia.
  3. dNTPS - Yes, required for all PCR reactions.
  4. primers for the insect gene - Yes, because they need to amplify the target insect sequence if that's the focus.
  5. DNA polymerase - Essential for the reaction.
  6. primers for the Wolbachia gene - Yes, if we want to amplify Wolbachia specifically.

Correct Answers:

  • Template DNA from an insect infected with Wolbachia
  • dNTPS
  • Primers for the insect gene
  • DNA polymerase
  • Primers for the Wolbachia gene

Question 23: Negative Control Tube of PCR Activity

In a negative control tube, it typically contains all reagents except for the template DNA, ensuring that any amplification is due to contamination:

  1. DNA polymerase - Yes, present in all controls.
  2. primers for the insect gene - Yes, should be included.
  3. template DNA from an insect infected with Wolbachia - No, absent to confirm no amplification due to contamination.
  4. primers for the Wolbachia gene - Yes, should be included.
  5. dNTPS - Yes, also required for the reaction.
  6. template DNA from an insect not infected with Wolbachia - No, as the template is absent in negative controls.

Correct Answers:

  • DNA polymerase
  • Primers for the insect gene
  • Primers for the Wolbachia gene
  • dNTPS

Question 24: NTC Tube of PCR Activity

In a Non-template control (NTC) tube, it typically contains all reagents except template DNA:

  1. template DNA from an insect infected with Wolbachia - Absent in NTC.
  2. template DNA from an insect not infected with Wolbachia - Absent in NTC.
  3. primers for the Wolbachia gene - Yes, should be included.
  4. dNTPS - Yes, should be included.
  5. DNA polymerase - Yes, should be included.
  6. primers for the insect gene - Yes, should be included.

Correct Answers:

  • Primers for the Wolbachia gene
  • dNTPS
  • DNA polymerase
  • Primers for the insect gene

Overall, always make sure to confirm the exact protocols you used for your PCR experiment to avoid incorrect selections.