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.