Team:EPF-Lausanne/Notebook/11 July 2012

From 2012.igem.org


Morning

The two plates, with RO + Spectinomycin, with 50 and 100 µg/ml, wrapped with parafilm and put in the fridge after overnight at 37ºC.

Preparing a Maxiprep for 12th July

Two maxipreps to be done:

  1. LovTAP + Ampicillin, transformed with our cells the 28 June.
  2. RO + Spectinomycin 100 µg/ml, transformed with our cells the 10 July.

We use 2 big erlenmeyers (like 1l), to allow the liquid to be shaken (bacteria like it).

We want a volume of 200 ml for each, with 100 µg/ml of antibiotic. This means 200 µl of antibiotic per maxiprep (antibiotics have both a concentration of 100 mg/ml).

Steps:

  1. Take the 2 plates from the fridge and let warm in oven for 5 minutes, 37ºC.
  2. Take the antibiotics from the freezer and let thaw.
  3. Fill the 2 erlenmeyers with 200 ml of LB each.
  4. Add 200 µl of antibiotic to each, spect to RO and amp to LovTAP.
  5. Pick a colony from each plate with a pipete tip and add it to the resp. erlenmeyer.
  6. Leave overnight in the incubator, turning, at 37ºC.


Protocol: None

Forgot to insert protocol.