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

 

B. Agrobacterium transformation with binaries


C. Antibiotic selection

  • Agrobacterium (GV3101)

Rifampicin 100 µg/ml

   
  • Ti-plasmid (pMP90RK)

Gentamycin 50 µg/ml,
Kanamycin 50 µg/ml

   
  • Binary (pLH vectors)

Spectinomycin 100 µg/ml,
Streptomycin 300 µg/ml