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12 Jun 2019

Agarose gel electrophoresis is used to separate charged molecules. Why do nucleic acids migrate from the cathode (-ve electrode) to the anode (+ve electrode) during agarose gel electrophoresis?

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Casey DurganLv2

12 Jun 2019

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in our lab, we used agarose gel electrophoresis to separate DNAfragments generated from the restriction enzyme digests of plasmidDNA.
Agarose can also be modified so that RNA can be separated. describethis modification.

1. On an electrophoresis power source the anode (positive charge) is red and cathode (negative charge) is black. When you put your gel in the rig, do you want the end with the comb/wells to be at the red or black end? Why?

2. The source of drag on a linear DNA molecule is the:

a. TAE Buffer

b. Nitrogen bases in the DNA molecule

c. Electric Field

d. Agarose

3. During agarose gel electrophoresis, the cause of drag on a linear DNA molecule is the:

a. Interaction with TAE buffer

b. Nitrogen bases in the DNA molecule

c. Interaction with Ethidium Bromide in the gel

d. Agarose in the gel

4. As part of your prelab work, you should have looked at the DNA isolation protocol. Why do you boil your tubes?

a. To disrupt the cell and nuclear envelope to release the DNA

b. To seperate the DNA from other cellular debris

c. To denature the DNA for PCR, the next step

d. To isolate cheek cells

5. At the end of your DNA isolation procedure you will have Chelex beads and some liquid. Explain which layer has your DNA (and thus you will save) and why.

6. What is the electric charge (q) on a 200bp linear piece of DNA?

rosecaribou448

ANSWER ALL

CELL AND MOLECULAR

1 When running DNA samples, the comb should ideally be placed?

A) in the gel near the positive electrode of the electrophoresis

B) in the middle of the gel equidistant from both electrodes

C) in the gel near the negative electrode of the electrophoresis chamber

2 How is agarose powder made to dissolve in the buffer?

A) hot plate

B) sonication

C) microwave

D) stirring

3 When the power supply is turned on, how can you immediately tell that the gel is running?

A) electrical discharge can be observed on the electrodes.

B) Bubbles form on the electrodes.

C) The DNA begins to move through the gel.

D) The gel begins to heat up.

4 Which of the following is NOT a purpose of the loading buffer used in our laboratory?

A) It helps to visualize the DNA bands after electrophoresis is complete.

B) It contains EDTA which uninhibited nucleases.

C) It is thick and viscous allowing the DNA sample to load into the well more easily.

D) It contains a dye that makes the DNA sample easier to see.

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Agarose gel electrophoresis is used to separate charged molecules. Why do nucleic acids migrate from the cathode (-ve electrode) to the anode (+ve electrode) during agarose gel electrophoresis?

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