How to make 500ml of 1% Agarose Gel Stocks.
Ingredients:
 5 g Agarose (Fisher BP1356)
 10 mL 50x TAE (242g Tris Base, 57.1ml Glacial Acetic Acid, 18.6g EDTA, MilliQ
water to 1L)
 MilliQ Water
 25 uL Ethidium Bromide (1% solution; Fisher BP1302)
Directions:
1) Measure out 5 g of Agarose (or 1g/100mL of solution) and place in graduated
cylinder.
2) Measure out 10 mL of 50x TAE (or 2 mL/ 100 mL of solution) and add to the
graduated cylinder.
3) Fill graduated cylinder with 500 mL MilliQ water.
4) Parafilm the top and mix thoroughly by inversion.
5) Immediately pour into a 500 mL bottle.
6) If any residual agarose is at the bottom of the graduated cylinder, pour some of the
TAE solution from the bottle back into the cylinder, swirl a few times, and pour back
into the bottle. Repeat until the cylinder has no trace agarose.
7) Lightly cap the bottle (such that steam can still escape) and microwave until the
solution becomes clear (~3-5 minutes). Make sure not to put it in for too long so the
water boils off.
8) Once the solution has cooled (you should be able to touch it for 10 seconds) add 25
uL of Ethidium Bromide solution using gloves (!) (or 5 uL per 100 mL solution) to
the bottle and mix. Final concentration if ethidium bromide is 0.5ug/ml.
9) Allow to cool to room temperature before pouring into gel mold.
Anjali Bisaria
6/24/2011