Team:TU Darmstadt/Notebook/Methods/Heat shock transformation
From 2014.igem.org
Heat shock transformation
Equipment:
- Heating bath
- Incubator
Chemicals & consumables:
- Ice
- Pipets + steril tips
- LB medium
- LB-Agar-Plates + antibiotics
Procedure:
- Defrost stocks of competent cells (100 µL in 1.5 ml Eppendorf tube) on ice.
- Add the DNA and incubate the suspension for 15 minutes on ice.
- Heat shock is done by incubating the cells for 45 seconds at 42°C. Put samples back on ice for 2 min.
- Add 1 mL of LB medium and incubate for 1 hour at 37°C in order to obtain antibiotic resistance.
- It might be useful to spin down cells at 5000 rpm for 5 min. Resuspend pellet in 100 µL LB.
Finally the cells are ready to be spread out.