Gel electrophoresis

Setting up and running of a DNA gel using electrophoresis.

[region]

Thank you very much for the visual explanation! I couldn't quite picture my upcoming lab until I watched your video. You did a phenomenal job of explaining the procedure!

[gkpeter]

The two bands are not DNA, but a chemical marker so you can monitor the process. DNA is clear, so you can't see it when in the gel until you use something to stain it.

[CiaoBello]

I once accidentally destroyed the agarose gel filling in my last sample, simply because i punctured the gel with the tip. Whats even funny is that i had a group of 5 others who were watching me as if their life depended on my hands.

[edyoucate]

Wow nice video, thanks for helping me understand this topic

[floranataline]

actually, the dyes stain the DNA to determined its position. yeah, DNA fragments carry a small negative charge (bcoz of the phosphate group) so DNA is attracted to the anode (the positive electrode) :)

[CathDamienn]

unintentional asmr ngl, was a very well made and informational video!!

[gkpeter]

Good question. The point is that to make a library, the vector can only accept certain size inserts. Therefore, you can use electrophoresis to select the size of the insert to increase efficiency.

[AskScience]

Hey great video.
I would have liked if you spoke about each of the material you used.

Ie. what the agrose is for, what the buffer is for and why we used it, what the loading dye's purpose is, etc.

[kedaovar]

i have a gel electrophoresis exam tomorrow, HATE this chapter, but you made it interesting

[gkpeter]

@ZScarabello There are lots of reasons to know the sizes. Verification of constructs, genotyping, cloning, etc. But the simple answer is, yes, you can extract the DNA from the gel if needed. I have done it several times.

[gkpeter]

@1006Will You can monitor it by keeping track of the migration dyes. In the video, you will see there are two colored dyes. The fastest moving one runs at about 500 base pairs. So, if you know the sizes of your DNA, you can prevent it from running into the next set of wells.
This whole thing can be done in 1.5-2 hours. Depends on your conditions and number of samples.

[HawaiiZach]

@gkpeter One reason I perform gel electrophoresis to identify the size of the DNA band is to determine if the primers I used in PCR (Polymerase chain reaction - used to amplify small quantities of DNA into the thousands) actually cut the DNA to the length I needed. If you are only focusing on looking at a certain section of DNA primers will "cut" the section out you need, then they will be amplified to the thousands using PCR. You can then run the product out on a gel to determine if it cut

[madgoldnz]

Good onya for making this, there is nothing like seeing it for real.

[phitox]

@ZScarabello actually not always the size is being determined. this is the way we can actually see if we have an insert (desired trait) in your DNA sample, and in this process we can cut the specific characteristic of your desired trait to put it in a transgenic species, it is important in Biotechnology, the result is called chimera. yes you can extract the DNA after separation..

[gkpeter]

@Peachpassion9 Compare it to a DNA ladder or molecular weight marker. You can guesstimate it or graph the known mass vs. length of migration from the well. You graph it on semi-log graph paper.

[gkpeter]

You choose the loading buffer based on the volume of your sample. If I have a 10x loading buffer and a 20 microliter same I will add 2 microliters.

[fcdog]

Thank you for the video! I have to do this in college soon. It was nice finding an electrophoresis video not in spanish finally hah

[gotb]

Bio 132 is the best class I've ever taken!

[jub]

Nice video! I have a cuestion
If I agarose gel 1%, TAE 1Xand small gel
How many volts I will use to running the electrophoresis?
How long (minutes) I is necesary to running the electrophoresis?
* I have extracted DNA of 500-700 pb

[Ranyaayoubi]

Great video! But what is the purpose of the TEA buffer, do you mind explaining?