SURE 2 Supercompetent Cell Transformation

(Stratagene Cat. # 200152)
updated 02/19/06 - Benjamin Costantino

(For the PDF version, click here.)

NZY+ Broth*
22g NZCYM Broth (Gibco cat. #13655022)
add deionized water until 1L
adjust pH to 7.5 and autoclave

1. Retrieve 100µl aliquot tube of Sure 2 cells from -80°C freezer and ß-ME provided by Stratagene. Allow the cells to thaw on ice.

2. Add 2µl of ß-ME to cells and gently swirl. Place on ice for 10 min, gently swirling every 2 min.

3. Add 5µl 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 30 sec.

5. Incubate the tube on ice for 2 min.

6. Add 900µl of preheated NZY+ broth and incubate tubes at 37°C for 1 hour, rotating.

7. Plate between 100-200µl of transformation mixture on B-AMP plates.

8. Incubate the plates overnight at 37°C.

* Prior to use, add the following filter-sterilized supplements:

12.5ml of 1M MgSO₄
12.5ml of 1M MgCl₂
10ml of 2M Glucose