How to Make and Run an Agarose Gel (DNA Electrophoresis)

Working with DNA in your lab? Well then here's a video for you. To check whether your PCR worked, or if you have an insert, you need to run a DNA gel.

Here Dan shows you how to make the agarose gel, and how to load and run your samples as well.

[lollypoppalace]

thank you so much!!! I remember the lab instructor expecting us to do this without any prior knowledge or instructions needless to say it was a tough day

[labtricks]

The purpose of using UV light is to see the bands on the gel. When you stain using ethidium bromide you can't see the bands until you shine UV light on it. Ethidium bromide reacts with the DNA in your gel by inserting into the DNA molecule. In the presence of UV light, ethidium bromide fluoresces and you can see where the DNA bands are located on the gel.

[labtricks]

@088zorba

-Wait for the gel to cool because it's really hot. Also, you don't want to create ethidium bromide (EtBr) vapours, so if you add EtBr when the gel is cool, you can prevent that.

-Add EtBr when the gel cools down

-When you take it out of the microwave, look at the solution and make sure that all of the agarose has been dissolved, and that it is a clear liquid. If you see gel like particles floating in it, microwave a little longer.

[labtricks]

In most cases the water loss is negligible (ie. if you are simply running a gel to see the presence of a band and the resolution or % agarose is not important). However, if you need to be very specific about your % agarose (i.e. you need exactly 1% agarose) then prior to microwaving you should weigh the flask (containing agarose and buffer). Then after microwaving when the agarose is dissolved, re-weigh and add water (not buffer) until you reach that same mass.

[labtricks]

@osehie Everyone uses different stains so we didn't cover that in this video. EtBr is a commonly used one and can be added either to the gel mixture or used for staining afterwards. That depends on personal preference.

[labtricks]

@088zorba

*Another way to stain using EtBr is to soak the gel AFTER running electrophoresis in a container of EtBr solution. This method prevents contamination of the gel electrophoresis equipment, and allows EtBr to be contained to one section of your lab. Also, no EtBr vapours :)

[SunilBirju]

Really a good video. Very informative indeed. Has anybody else tried loading samples outside the tank and then putting it in.

[elliottstokes]

Thanks for helping in my competency test :)

[gufranali]

Thanks alot. I really need this video to share it with my students.

[KosbyShow]

Thank you for these videos. They are helping me with this job interview

[orangensaftization]

great video! I'll have to do it tomorrow in my university lab course

[Afiqsenen]

Thnx for uploading this video..it's really help me...

[terrylaitw]

Great video, very concise and straight forward. are you from Taiwan??!

[labtricks]

@beeryya You can use TAE as well, it's up to you and based on your protocol. If your protocol specifically asks for TAE then use that. Otherwise TBE works just fine!

[tanimbmb]

Thank you very waiting to watch the video.

[FereldenRefugee]

thanks!!! this really helped me understand gel electrophoresis, which is good cuz I'm doing a science project on it!!! :)

[ChemistDrummer]

You could make a video for the visualisation process with EtBr also...

[labtricks]

Could you specify which calculations you need help with? For making an agarose gel, the calculation needed is pretty much what is shown at 0:19 of this video. Are there any other calculations you guys need help with?

[labtricks]

Yes we are working on another video for that. In the meantime, if you need help with that part of the protocol, feel free to ask us questions :)

[eerielconstantine]

omg thank you for this, I can impress my potential future job XD