Agarose gel electrophoresis , 1.2 % with Ethidum bromide
Weigh out 0.6 g ( 1.2 % w / v of 50mL ) agarose and add it to the Erlenmeyer Flask .
Add 50mL of 1x TAE buffer Place Erlenmeyer Flask in microwave .
Set to wait 30 seconds , then full power ( P10 , 1250W ) for 20 seconds followed by low power ( P1 , 125W ) for 30 seconds or until solution is clear and agarose is completely dissolved .
Remove Erlenmeyer Flask from microwave and let it sit on the lab bench to cool just until you can comfortably pick it up .
Add 1μL concentrated ethidium bromide ( 10mg / mL ) into the flask and swirl to mix , taking care not to introduce bubbles .
Place gel tray on clamp and clamp securely .
Add well plates where you want wells and use a level to ensure it is balanced .
Pour contents of the Erlemeyer Flask into the gel tray and let it sit for 30 minutes , or until a blue tint appears .
Remove the well plates carefully as to not tear the gel and remove the tray from the clamp , but ensure the gel remains in the tray .
Place gel tray into gel electrophoresis apparatus with the wells closer to the negative / black end .
Pour additional TAE Buffer to fill each side of the apparatus and to create a thin layer of buffer covering the top of the gel .
Prepare DNA ladder and samples by adding 6x blue dye Pipette your samples into each well .
Place lid on apparatus and plug cables into high voltage power supply .
Run at 100V ( ~ 6.6V / cm ) for 45 - 60 minutes or until the loading dye has sufficiently migrated down the gel .
Gel can be imaged on UV transilluminator through the UV - transparent gel tray or removed and wrapped in plastic wrap for storage at 4°C for later use .
