Journal Club
Light Regulation of Plant Growth and Development
Building LED Light Sources for Plant Research
Spectra of Light Sources Used in Published Experiments
Strawberry Transformation
Making Transgenics using the Laboratory Festival #9 Line
1. Early day 1. Pick a fresh colony of Agrobacterium hosting the construct of interest, grow it overnight in 3 ml of LB with proper selection at 28°C.
2. Day 2. After approximately 24 h transfer the 3 ml to 50 ml LB with selection. Grow to an OD of 0.5 at 28°C.
3. Spin down cells 10 min in a sterile 50 ml conical tube at 1000 xg (~2800 rpm in our centrifuge).
4. Resuspend cells in co-cultivation medium to an OD of 0.5 containing 0.1 mM acetyosyringone (add 19.62 mg of AS to 400 ul of 75% EtOH. This is 0.250 M. Add 0.4 microliters per ml of co-cultivation media to give a final concentration of 0.1 mM, e.g. 10 ul per 25 ml).
5. Cut explants into agrobacterium suspension. Co-cultivate for 1 h at 23°C.
6. Pour off excess culture and place explants onto media 7 with timentin (or carb/cef).
7. Incubate in darkness for two days, move to light conditions.
8. After one week move the explants to 7 media with timentin and Kan or Phos.
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