1. Thaw on ice one tube of competent bugs.
2. Take 1/2 of each ligation into 1.5 ml tubes &
freeze reminder @ -20 C.
3. Take 10 µl of positive control KS+ into 1.5ml tube (control).
4. Add 100 µl of competent bugs to each tube.
5. Incubate 1 hour on ice.
6. Heat shock 5 min @ 37 C.
7. Add 1ml of LB medium to each tube.
8. Incubate @ 37 C water bath (no shaking) for 1 hr.
9. Dry plates 15' @ 37 C before inoculation.
10. Plate 200 µl from each tube (+20 µl Amp)
11. Spin down tube (with insert) @ 14 k for 30''.
12. Remove 700 µl supernatant.
13. Resuspend bugs & inoculate another plate.
14. Incubate plates @ 37 C ON.