Team:Groningen/Protocols for Lactococcus
From 2010.igem.org
(Difference between revisions)
Neima (Talk | contribs)
(New page: '''Preparation of cells''' *10 ml ON culture (SMGG) in 100 ml SMGG *Grow to OD600=0.2-0.7 *Wash three times with 50 ml icecold wash buffer*Resuspend in 1 ml wash buffer '''Electr...)
Newer edit →
(New page: '''Preparation of cells''' *10 ml ON culture (SMGG) in 100 ml SMGG *Grow to OD600=0.2-0.7 *Wash three times with 50 ml icecold wash buffer*Resuspend in 1 ml wash buffer '''Electr...)
Newer edit →
Revision as of 20:49, 27 October 2010
Preparation of cells
- 10 ml ON culture (SMGG) in 100 ml SMGG
- Grow to OD600=0.2-0.7
- Wash three times with 50 ml icecold wash buffer*Resuspend in 1 ml wash buffer
Electroporation
- 1 μl DNA in 40 μl cell-suspension in ice cold cuvette
- Electroporate at 2.5 kV, 25 μF, 200 Ohm.
- Add 4 ml SMG17MC*Incubate 2 hours (Ery induction=50 ng/ml)
- Concentrate cells to 0.5 ml.
- Plate on GSM17- Agar.