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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.