pBI121-MAP30

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	Leaf explants of 1–2 cm were inoculated for 5 min with an A. rhizogenes culture that had been grown overnight (to OD600 nm = 0.5), and then these materials were co-cultured on solid MS medium at 25°C in the dark.

Protocol tips
Leaf explants of 1–2 cm were inoculated for 5 min with an A. rhizogenes culture that had been grown overnight (to OD600 nm = 0.5), and then these materials were co-cultured on solid MS medium at 25°C in the dark.

Publication protocol

"The plasmid was extracted from transformed E. coli using a Plasmid Miniprep Kit (Fermentas). The recombinant plasmid was diluted 10 times, and 10 μL of this sample was used to transform 100 μL of A. rhizogenes strain ATCC AR15834 (at OD600 nm = 1) using the freeze-thaw method. One milliliter of liquid LB was added, and the cells were incubated at 28°C in the dark for 2 h. Then, the transformed bacteria were dispersed on LB agar containing kanamycin and rifampicin (100 mg/L) and were incubated at 28°C in the dark for 48 h. Colonies were confirmed to be recombinant using a specific PCR assay and by digestion of the extracted plasmid.
Leaf explants of 1–2 cm were inoculated for 5 min with an A. rhizogenes culture that had been grown overnight (to OD600 nm = 0.5), and then these materials were co-cultured on solid MS medium at 25°C in the dark. After three days, the explants were transferred to fresh MS medium supplemented with 400 mg/L of cefotaxime and 150 mg/L of kanamycin and were maintained at 25°C under a 16/8 h light/dark photoperiod for two weeks. The hairy roots that formed at the incision sites of the leaf fragments were subsequently transferred at two-weeks intervals to fresh MS agar containing cefotaxime and kanamycin at the concentrations noted above and were incubated at 25°C in the dark."

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