McGill/Plating

From 2007.igem.org


Ingredients:

1) LB - 15.5g LB to 500ml dH20
2) Agar - 6.0g Agar to 200ml dH20
3) Amp & Kan antibiotics.

Then Autoclave both bottles.

  • If the Agar is already prepared, use a microwave to melt the agar (remember to make sure the top of the bottle is loosely screwed).


Procedure:

  1. Mix together the LB and the Agar into a single bottle. If the Agar has been microwaved, allow it to cool to <60C before mixing.
  2. Swirl the new solution well.
  3. Add the antibiotics.
    1. For Amp: 1ul/ml. eg. for 700ml LB+Agar soln add 700ul.
    2. For Kan: 10ul/ml. eg. for 700ml LB+Agar soln add 7ml.
  4. Spread evenly on plates, making sure there are as few bubbles as possible.



NOTE: Make sure when stacking the new plates, to stack them with the culture on the top and the lid of the plate at the bottom. This is to prevent any water accumulation from the fridge or condensation dripping onto the medium.