IGEM:Harvard/2010/General Protocols/Agrobacterium

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We have received from Amy Hark the agrobacterium strain: GV3101 pMP90. This is the strain that was used to transform the plant vectors we purchased into Arabidopsis.

  • Frequently used for many binary vectors for Arabidopsis
  • Resistance to Rif (10mg/L) (in genome), Gent (30mg/L) (in helper plasmid), and possibly Kan (30mg/L) (in helper plasmid) ---- she told me she usually only selects with Gent, but we may want to try and see if it is still resistant to Kan
  • Known to develop spontaneous Tet resistance, so this strain should be used with care.
  • Fast growing colonies after transformation are false. True transformants appear 1-2 days after fast growing colonies.
  • Use 100ug/mL timentin/cefotaxim for Agro removal when selecting transgenic plants on plate
  • Use bromin to kill agro.