Gel Electrophoresis Protocol

Summary

This video script outlines the assembly and operation of a gel electrophoresis system. It details the setup process, including connecting components like the camera hood, power supply, and laptop. The script emphasizes the importance of proper connections and initial checks before conducting experiments. It also instructs on preparing pre-cast gels, loading samples, and running the gel electrophoresis. The video demonstrates how to capture and save images of the gel for analysis, providing a comprehensive guide for beginners in DNA analysis.

Takeaways

• 🔌 **Power Connection**: Connect the power supply to a wall power outlet and ensure the laptop is plugged in due to its weak battery.
• 💻 **Laptop Setup**: Log in to the laptop using the provided username and password, and confirm it's powered on.
• 📸 **Camera Hood**: Connect the camera hood directly to a USB port on the laptop without needing extra power.
• 🧪 **Gel Preparation**: Remove the packaging from the gel cassette and ensure it fits properly into the gel box.
• 🚿 **Cleaning Wells**: Wash the wells with water using a squirt bottle, then dry with a Kim wipe or paper towel.
• 🔬 **Sample Loading**: Load samples into designated wells, avoiding bubbles and ensuring they settle properly.
• 📊 **DNA Ladder**: Add the DNA ladder to the first well of each row, following the same loading technique as for samples.
• 🔌 **Running the Gel**: Set the power supply to 275 volts and run for the specified time, ensuring the cords are connected correctly.
• 👀 **Visualizing DNA**: Use the blue light on the gel box to visualize the DNA bands migrating through the gel.
• 📸 **Capturing Images**: Use the camera hood to capture images of the gel and save them with recognizable names and locations.
• 🔄 **Post-Experiment**: Turn off the power supply, unplug cords, and transfer images to personal storage devices if needed.

Q & A

What components are necessary to assemble the Lonza system?

The necessary components include a camera hood, a power supply, a gel box, a laptop, and cords.

What color are the cords that need to be connected to the power supply?

The cords are red and black.

How should the power supply be connected to the wall outlet?

The power supply should be connected to the wall outlet using the provided cords, but only right before running the gel.

What is the purpose of the camera hood?

The camera hood should be plugged directly into a USB port on the laptop and does not require an extra power supply.

Why should the laptop be plugged into a wall power outlet?

The laptop should be plugged into a wall power outlet because the battery on these repurposed laptops has little to no life.

What is the purpose of the pre-cast gel?

The pre-cast gel is used to visualize samples in an experiment and is appropriate for the class size and sample needs.

How should the wells of the gel be prepared before loading samples?

The wells must be washed with water, excess water should be poured out, and the gel dabbed with a Kim wipe or paper towel before returning it to the gel box.

What is the significance of the provided DNA ladder?

The DNA ladder is provided for visual size comparison of the samples.

What is the correct way to load a sample into the wells of the gel?

Three microliters of the sample should be loaded into its designated well, avoiding bubbles or flow out of the well.

Why should the first well of the gel be skipped when loading samples?

The first well is reserved for loading the ladder, which will be done last.

How long should the gel be run at 275 volts?

For a one-tier gel, it needs to have electrical current for 4 minutes.

How can you capture an image of the gel after the electrophoresis is complete?

After the electrophoresis is complete, press the camera icon on the computer to capture the image and save it in a recognizable location with a recognizable name.