E. coli transformation
1. Thaw frozen (-80 ° C) competent cells on ice
2. Mix 1 µl DNA
3. Incubate on ice 20 min
4. Incubate 42 ° C 45 sec-1 min
5. Incubate on ice 2 min
6. Add 1 ml LB
7. Incubate at 37 ° C 45 min-1hour
8. Spin cells down (2min at 14000rpm)
9. Remove supernatant, keep cell pellet in a small amount of medium (about 50-100 µl.
10. Re-suspend cells in the medium, transfer everything on LB agar plate (+antibiotic) and spread cells with a blue sterile spreader. (The goal is to evenly distribute cells and to allow them to be absorbed into the agar).
LB ((+antibiotic) agar plate:
1. Heat 150 ml LA agar in a microwave oven (9 –for 3 min on old microwave or power 5 for 3 min on new) until completely melted.
2. After cooling the solution and add antibiotic
3. Make Petri dishes (150 ml LA makes 5-6 plates)