Prep: Make stock solution of 1.00x TAE buffer for use in gel casting and buffer solution in gel boxes during electrophoresis.
- Using a graduated cylinder, measure out 100mL of 10x TAE buffer.
- Using a graduated cylinder, measure out 900mL of DI H2O.
- Combine the two volumes in a 1000mL orange-capped Pyrex jar.
- Label jar with TAE concentration, your initials, and the date.
Directions for 4" x 5.75" gel (used in gel extraction):
- Set up gel mold with comb in place. MAKE SURE THE TRAY IS LOCKED IN PLACE. Use bubble-level to ensure even settling of gel matrix.
- Add 2.25g agarose to a 500mL Erlenmeyer flask
- Add 225mL of 1.00x TAE buffer to the flask
- Microwave flask for 2 minutes
- After 2 minutes make sure agarose is completely dissolved in TAE. Swirl flask if it is not, microwave for another 15-20s if needed.
- let it cool for 2-3 min, but make sure it does not solidify.
- Add 2μL ethidium bromide to flask.
- Pour into gel mold with comb in place.
- Let sit for ~30 minutes to 1 hour.
- While allowing gel to set, fill the 500mL Erlenmeyer flask with ~300mL of water
- microwave for 2.5 minutes.
- swirl and empty into drain.