Oct 18, 2020
Agarose Gel Electrophoresis
- 1South China University of Technology
Protocol Citation: Jiaxin Li 2020. Agarose Gel Electrophoresis. protocols.io https://dx.doi.org/10.17504/protocols.io.bhvpj65n
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: June 24, 2020
Last Modified: October 18, 2020
Protocol Integer ID: 38543
Mix
Mix
Preparation of TAE
Preparation method:
1. Concentration: 50x to 1x
2. Preparation:
A) 100ml 50x mother liquor to beaker, Gatrochen water to 500ml
B) Open a new tank of 4.5L Watson's water, pour the above liquid into 5L liquid
Prepare 1% agarose (Commonly used for 200 bp-5 Kb of DNA) :
| agarose/g | TAE/ml | dyestuff/ul | hole count | |
| 0.8 | 80 | 2 | 50 | |
| 0.4 | 40 | 1 | 25 | |
| 0.2 | 20 | 0.5 | 11 |
Process
Process
Melt agarose in 1X TAE buffer in microwave oven until the liquid is fully transparent.
Add EB (Ethidium bromide) in the melted agarose.
Pour the melted agarose in the gel cast with the comb set.
Wait 25 minutes until the gel solidifies.
Cover the gel with 1X TAE buffer and remove the combs carefully.
Load the samples in the wells:
3 μl of 1 kb DNA ladder
Mix 3 μl of DNA with 1 μl loading buffer
Run the gel at 120 volts for 25 minutes without letting the bands run out of the gel.
Remove the gel from the chamber.
Visualize the DNA fragments.