STRAINS:

Helper - HB101 pRK2013, kan^r
Donor - DH5a pLAFR3, tet^r

^METHOD:

1. Grow cells overnight.
2. Dilute 100 µl of each culture into 5 ml LB.
3. Filter onto sterile 0.2 µM filter.
4. Place filter onto plate of LB; leave overnight.
5. Place filter in tube with 1 ml water and vortex.
6. Plate onto minimal media, 1, 10, and 100 µl.
7. Incubate for 1-2 days.