Difference between revisions of "Jones Lab:Protocols:Agrobacteria"

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(New page: <h3><font style="color:#F8B603;">Competent Agrobacteria (for cold-shock transformation)</font></h3> ---- #O/N starter culture (with appropriate antibiotics; GV3101 should be grown in Rif a...)
 
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Latest revision as of 03:37, 29 October 2012

Competent Agrobacteria (for cold-shock transformation)


  1. O/N starter culture (with appropriate antibiotics; GV3101 should be grown in Rif and Gent), at 28°C
  2. Inoculate 500ml LB with starter culture
  3. Grow at 28°C until OD 0.5-1.0
  4. Chill culture on ice for 30 min
  5. Spin at 10k for 15 min in SS34 at 4°C
  6. Wash in 50 mL ice-cold 20mM CaCl2
  7. Spin at 10k for 15 min in SS34 at 4°C
  8. Resuspend in 10 mL ice-cold 20mM CaCl2
  9. 100 μL aliquots, flash-frozen in N2