Direct Agrobacterium transformation of GV3101(pMP90RK) with pLH vectors
Although the transformation frequency by this method is low compared to the triparental mating method, the technique is reliable and very rapid. This transformation procedure also eliminates much of the plasmid rearrangement that often occurs during triparental mating.
A. Preparation of competent cells
Transformation of E. coli
- Thaw competent cells carefully on ice
- Incubate the DNA with the competent cells for 1-10 min on ice
- Incubate for 30-90 sec. at 42°C - heat shock -