How to Make and Run an Agarose Gel (DNA Electrophoresis)
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- čas přidán 17. 07. 2024
- 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.
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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
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 :)
Thanks alot. I really need this video to share it with my students.
@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.
Thank you for these videos. They are helping me with this job interview
Really a good video. Very informative indeed. Has anybody else tried loading samples outside the tank and then putting it in.
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.
@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.
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.
Thanks for helping in my competency test :)
great video! I'll have to do it tomorrow in my university lab course
Thnx for uploading this video..it's really help me...
Thank you very much.....eagerly waiting to watch the video.
really informative and interesting
Very enjoyable thank you
thanks!!! this really helped me understand gel electrophoresis, which is good cuz I'm doing a science project on it!!! :)
it was useful, thanks
The camera we use is the Kodak Zi8.
Great video, very concise and straight forward. are you from Taiwan??!
@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 :)
omg thank you for this, I can impress my potential future job XD
Thank you!
You could make a video for the visualisation process with EtBr also...
Good one
Video on calculations that go into making such gel
czcams.com/video/Z--iWjaodmo/video.html
@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!
Yes! Email us at info@labtricks. com to find out how :)
Thank you very much. I've learned some techniques from your tutorial. But this video doesn't include the preparation of sample before loading onto the gel. Could you please add this?
I love your HD camera. What maker/model is it? Thanks.
how much loading dye should I add to the sample?
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?
@Saran89 Nice trick, thanks for sharing!
Very nice .
I have two questions.
1- I want to make a complete gel without having water on it. In other words I want it to be dry. what should i do?
2- What is the melting temperature of this gel that you made? I think it should be around 45c. How can I make a gel with much higher melting temperature?
it is very helpful. but you should also show what is happing after get result.hope for improve
What is the concentration of buffer to be used in gel tank? Will 1x suffice
my bacterial genomic DNA sample load well thereafter time migrate sample. migrated sample no DNA band , the load on well are presented DNA . what reason ?
Is it possible to buy these videos or dowload them?
Hi dont you loose some water due to evaporation while your are microwaving it ?
I am using the exact same electrophoresis system, but I have a couple of problems. The gel usually fills up behind the plastic that holds the gel. Also, the power supply heats up and starts blinking and the bands turn up very uglish.
1) After casting, the excess gel behind the plastic tray can easily be peeled off and discarded.
2) If the power supply is heating up and the light is blinking, the current has exceeded the limit of the system. Make sure your buffer is correct. If you are reusing buffer, the water evaporates and increases the salt concentration, which increases the current and heat. So it is recommended to discard the old buffer and replace with fresh 1X TBE buffer. If you want to keep reusing old buffer, then top off the old buffer with water and NOT with more buffer - remember you want to replace the water that's been evaporated to maintain the original salt concentration.
Hope that helps!
Dude I love your video, and forgive me I don’t mean to be offensive, but every time you say agarose I hear egg rolls😅 so I started busting a gut at the microwave egg rolls for two minutes part. Again great work!
Drew Parks I want one too
Why TBE? Why not TAE? (Tris/ Acetic acid/ EDTA)
Gel extraction for a TBE gel is different, but the gel doesn't heat as much I think. I use TAE.
1 ml of agarose = 1 g ?
Hmm....DNA Electrophoresis huh? Where's the Ethidium Bromide???????
Major_Lag I think last person who used EtBr died of some sort of cancer
gelred is common now. it´s less dangerous
Also except of gelred you can use methylene blue as an alternative dye to the fluorescent dye EtBr
Really a good video. Very informative indeed. Has anybody else tried loading samples outside the tank and then putting it in.
really informative and interesting