Begin by adding 50 microliters of electrocompetent Agrobacterium tumefaciens cells into a pre-chilled 0.1-millimeter Q-vet. Add two microliters of pCAMBIA1302 to the Q-vet. now use an electroporator at 2, 400 volts to eelectroporate the cells.
Immediately add one milliliter of LB media to the Q-vet and pipette the solution up and down to mix gently. Next, transfer the resuspended cells into a 1.5-milliliter microtube. Then incubate it at 24 to 30 degrees Celsius for a minimum of one hour to recover.
Now plate the cells on antibiotic supplemented LB auger. Incubate the plates at 28 to 30 degrees Celsius for two to three days. Finally, inoculate a single colony in LB broth.
Allow the culture to grow overnight. Then cryopreserve the plasmid-containing strain with a 50%glycerol solution at 80 degrees Celsius for future use.
