Arabidopsis floral dip transformation

Дата канвертавання18.04.2016
Памер3.29 Kb.

15 March 2007

Arabidopsis floral dip transformation

  1. grow plants at an optimal condition. Make sure they grow robustly.

  2. Decap each plant when it start to bolt. When the stems are about 5 cm, they are ready for transformation.

  3. Transform plasmid into agrobacterial competent cell using electroporation. Identify the right transfomants.

Day 1

Inculate a single colony in 5-10 ml LE selection medium and shake at 300 rpm at 28-30 C for 2 days
Day 2

Water plants

Day 3

  1. Transfer 5 ml culture into 500 ml LB selection medium and shake for overnight.

  2. Pellet cells at 5000 rpm for 10 min at 4C. Discard the supernatant.

  3. Resuspend cells in 1L of 5% sucrose solution + 10 mM MgCl2

  4. Add 500 l silvet (0.05%). Use a stir bar to mix well.

  5. Dip inflorescences of each plant into the ag-bacterium solution for 1 min

  6. let plants lie on the side in the tray.

  7. Cover the plants with another tray for 24h or overnight to avoid evaporation

  8. Let plants grow at normal condition.

База данных защищена авторским правом © 2016
звярнуцца да адміністрацыі

    Галоўная старонка