High Resolution Agarose Gel Electrophoresis
互联网
986
实验步骤
NOTE: Chill Agarose 1000 gels for 30 minutes at 4°C before use to achieve best resolution.
Method 1: Microwave (recommended for < 3% concentrations)
- Into a flask holding 2-4 times the desired solution volume, add a magnetic stir bar and the calculated volume of buffer at room temperature.
- Put the flask on a magnetic stirrer and slowly sprinkle the calculated amount of agarose powder into the flask while stirring constantly to prevent the formation of agarose clumps.
- Remove the stir bar
- Weigh the flask and solution before heating.
- Place in the microwave oven and heat on high power for two minutes.
- Remove carefully as any microwaved solution may become superheated and foam over when agitated. Gently swirl to resuspend any agarose particles.
- Reheat on high power using 15-20 second intervals or until the solution comes to a boil, and solution is complete.
- Remove carefully and gently swirl.
- Return the flask to its original weight by adding warm distilled water.
- Mix gently and cool to 50-60°C (at room temperature for at least 20 minutes) before pouring the solution into the tray.
- During the cooling time any air bubbles will disappear.
Dissolving Agarose 1000 (4%-5%)
Method 2: Boiling water bath (recommended for all concentrations, especially 4%-5%)
- Into a flask holding 2-4 times the desired solution volume, add a magnetic stir bar and the calculated volume of buffer at room temperature.
- Put the flask on a magnetic stirrer and slowly sprinkle the calculated amount of agarose powder while stirring constantly to prevent the formation of agarose clumps.
- Weigh the flask and solution before heating.
- Bring the solution to a boil while stirring and allow to gently boil for approximately 15-20 minutes or until the agarose dissolves completely.
- Return the flask to its original weight by adding warm distilled water.
- Mix gently and cool to 50-60°C (at room temperature for at least 20 minutes) before pouring the solution into the tray.
- During the cooling time any air bubbles will disappear.
Method 3: Autoclave (recommended for all concentrations, especially > 5%)
- Into a flask holding 2-4 times the desired solution volume, add a magnetic stir bar and calculated amount of buffer at room temperature.
- Put the flask on a magnetic stirrer and slowly sprinkle the calculated amount of agarose powder into the flask while stirring constantly to prevent the formation of agarose clumps.
- Heat two minutes in the microwave oven at medium power.
- Cover the opening of the flask with an aluminum foil to prevent spillover and autoclave at 121°C for 15 minutes.
- Remove from the autoclave and allow to cool to 50-60°C before pouring the solution into the tray.
