EDIT: Other transformation protocol has been updated by Brian. Follow that protocol.
SET UP
Make sure heat block is set to 42 C with water in the wells
Warm your plates and S.O.C to 37 C
Make sure the rest of your equipment is out on the lab bench and ready to use
You do not have time to gather equipment once your competent cells are out of the -80
PROCEDURE
- Bring the cells from the -80 and immediately onto ice
- Let them thaw for no more than 3-4 minutes
- Add in 1-2 uL of DNA and stiir
- Turn on the heat block to 42 C
- Wait 30-45 minnutes
- Before you use the S.O.C, make sure it's between room temperature and 37 degrees
- Heat shock DNA for 10-40 seconds
- Put it on ice for 2 minutes
- Add 450 uL of S.O.C
- Shake at 280 rpm and 37 C for 60 minutes
- Make sure your plates are already at 37 C
- Plate 100 uL or reaction and 10 uL of plasmid
- Incubate at 37 C for 16 hrs