Gel Electrophoresis Overview

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Questions and Answers

What does gel electrophoresis enable us to do?

Sort and measure DNA strands.

What is the filter that sorts the DNA strands?

The gel.

Where do we place DNA at one end of the gel?

In holes, or wells.

What do you add to make the DNA move?

Electrical current. Signup and view all the answers

What strands move quicker through the gel?

Short strands move quicker; long strands are slower. Signup and view all the answers

What process makes the sorted groups of DNA molecules visible?

Staining. Signup and view all the answers

What are the 5 steps used in gel electrophoresis?

<ol> <li>Make the gel 2. Set up the gel apparatus 3. Load the DNA sample into the gel 4. Hook up the electrical current and run the gel 5. Stain the gel and analyze the results.</li> </ol> Signup and view all the answers

What are the steps for making gel?

<ol> <li>Put a small amount of agarose into a flask 2. Add some liquid buffer to the flask 3. Place the flask into a microwave 4. Pour the melted agarose into the mold 5. Place the comb into one end of the gel 6. Let the gel cool and solidify 7. Remove the comb.</li> </ol> Signup and view all the answers

What do you need to set up the gel apparatus?

Gel, electrophoresis box, and buffer. Signup and view all the answers

What do you do to set up the gel apparatus?

Pour the buffer into the electrophoresis box and place the gel into the electrophoresis box. Signup and view all the answers

What are the steps for loading the DNA sample into the gel?

<ol> <li>Add loading dye to DNA 2. Suck up DNA into pipette tip 3. Eject DNA sample into well 4. Suck up DNA standard with new pipette tip 5. Transfer DNA standard into well.</li> </ol> Signup and view all the answers

How do you hook up the electrical current and run the gel?

Plug the black cord into the side closest to the wells and plug cords into the power supply, matching colors. Signup and view all the answers

How can you tell if a current is running?

Check for tiny air bubbles. Signup and view all the answers

DNA moves toward what charge and why?

Towards the positive charge because it is being repelled by the negative charge. Signup and view all the answers

What do you do to stain the gel and analyze your results?

Place the gel into the DNA staining solution (ethidium bromide), remove the gel after 30 minutes, place it on the UV light box, turn on the UV box while wearing protective gloves, and compare the bands from the DNA sample with the bands of known lengths. Signup and view all the answers

Study Notes

Gel Electrophoresis Overview

Gel electrophoresis is a technique used to sort and measure DNA strands by size.

Gel as the Sorting Filter

The gel serves as a filter that separates DNA strands based on their length.

Loading DNA into the Gel

DNA is placed into specific slots called wells at one end of the gel.

Movement of DNA

An electrical current is applied to the gel, causing the DNA to move.
Short DNA strands migrate faster through the gel compared to longer strands.

Visualization of DNA

Staining is the process used to make the sorted DNA molecules visible after separation.

Steps in Gel Electrophoresis

Five main steps are involved in the gel electrophoresis procedure:
- Prepare the gel.
- Set up the gel apparatus.
- Load DNA samples into the gel.
- Connect the electrical current and run the gel.
- Stain and analyze the gel results.

Gel Preparation Process

To prepare the gel, follow these seven steps:
- Combine agarose with a liquid buffer in a flask.
- Heat in a microwave until melted.
- Pour the melted mixture into a mold.
- Insert a comb to create wells.
- Allow the gel to cool and solidify.
- Remove the comb, leaving wells for the DNA samples.

Setting Up the Gel Apparatus

Essential components for the gel apparatus include:
- Gel, electrophoresis box, and buffer solution.
Setup involves pouring buffer into the box and placing the gel within it.

Loading DNA into the Gel

Steps to load DNA samples include:
- Mixing the DNA with loading dye.
- Using a pipette to transfer DNA into the wells.
- Adding DNA standards using a clean pipette tip.

Running the Gel

To initiate the gel run, connect the power supply:
- Black cord is connected to the side of the gel closest to the wells.
- Ensure proper connection by matching the colored cords.
Signs of current flow include the appearance of tiny air bubbles.

Charge Movement of DNA

DNA moves toward the positive charge because of its negative charge, being repelled by the negative pole.

Staining and Analyzing Results

To stain the gel:
- Submerge the gel in a DNA staining solution (e.g., ethidium bromide).
- After 30 minutes, view the gel under UV light.
- Protective gloves should be worn during the UV exposure.
Compare the stained bands with known DNA length standards for analysis.

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Description

This quiz provides an insightful overview of the gel electrophoresis technique used for separating and measuring DNA strands by size. It covers the process of loading DNA, the physics behind DNA movement, and the visualization methods following separation. Perfect for students studying biology, genetics, or molecular biology.