Summary: Benjamin Costantino-2/19/06
SURE 2 Supercompetent Cell
(Stratagene Cat. # 200152)
1. Retrieve 100ul aliquot tube of Sure 2 cells from -80°C freezer and _-ME provided
by Stratagene allow the cells to thaw on ice.
2. Add 2ul of _-ME to cells and gently swirl. Place on ice for 10min., gently swirling
every 2 min.
3. Add 5ul of ligation sample to the tube, gently swirl and place on ice for 30min.
(total conc. of DNA should be .1-50ng for max. efficiency)
4. Heat pulse the tube in heating block for 30sec.
5. Incubate the tube on ice for 2min.
6. Add 900ul of preheated NZY+ broth and incubate tubes at 37°C for 1 hr. rotating.
7. Plate between 100-200ul of transformation mixture on B-AMP plates.
8. Incubate the plates overnight at 37°C
22g NZCYM Broth (Gibco cat. #13655022)
ADD deionized H2O until 1 L
Adjust pH to 7.5 and autoclave