Box 1. Transformation of Agrobacterium
From the following article
Imogen A Sparkes, John Runions, Anne Kearns & Chris Hawes
Nature Protocols 1, 2019 - 2025 (2006) Published online: 30 November 2006
doi:10.1038/nprot.2006.286
This method is based on a freeze/thaw protocol20.
(i) Add 25 ng DNA per 50 
l of competent cells to 50 l of frozen, heat-shock competent bacteria in a 1.5-ml Eppendorf tube. It is usually sufficient to just add 5 l of miniprep DNA to the frozen bacteria.
(ii) Let cells thaw on ice for 10–15 min.
(iii) Freeze tubes in liquid nitrogen.
(iv) Heat shock in a water bath at 37 °C for exactly 5 min directly from the liquid nitrogen.
(v) Add 1 ml of YEB medium and incubate in an orbital shaker at 200 rpm, 28 °C for 2–4 h.
(vi) Centrifuge at 3,000 rpm at room temperature for 3 min, discard the supernatant and resuspend the pellet in 200 l YEB medium.
(vii) Spread on YEB 1.5% agar plates that contain the appropriate antibiotics (e.g., gentamycin, rifampicin and kanamycin) and incubate the plates at 28 °C for 36–48 h until colonies appear.
