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Agarose Gel and Running Buffer Recipes
Agarose Gel Recipes
- Agarose (see chart)
- 1x running buffer
- 0.1% EtBr (0.5mg/mL stock solution)
- Microwave for 1-3 minutes until agarose is dissolved, watching carefully as to not over boil solution
- Add EtBr after microwaving
- Make 100 mL of agarose solution for a typical cast
| % Agarose concentration | DNA resolution |
| 0.5 | 1– 30 kb |
| 0.7 | 800 – 1.2k bp |
| 1.0 | 500 – 10k bp |
| 1.2 | 400 – 7k bp |
| 1.5 | 200 – 3k bp |
| 2.0 | 100-2k bp |
DNA Running Buffers
10X TAE (for fragments >4kb)
- 48.4 g Tris base
- 11.4 mL glacial acetic acid
- 50 mM EDTA
- QS with H2O to 1L
10X TBE (For 0.1-3kb fragments and for higher voltages of >150V)
- 108 g Tris
- 55 g boric acid
- 9.3 g EDTA
- QS with H2O to 1L
Typically, gels can be run at 80-150V until dye line is most of the way through the gel. Take care that voltage isn’t too high or the gel will melt.