Team:UNIPV-Pavia/Methods Materials/Transformation

From 2009.igem.org

Revision as of 11:38, 30 June 2009 by Letizia (Talk | contribs)

EthanolPVanimation.gif



Protocols


Transformation

(estimated time: 3 hours and 30 min + 12-16 hours overnight incubation)

  • Put 1-2 µl of DNA resuspension or ligation into TOP10 tube.
  • Incubate on ice for 30 min.
  • Heat shock: 42°C for 1 min.
  • Put transformed TOP10 tube on ice and then add 250 µl SOC medium.
  • Incubate 1 hour at 37°C, 220 rpm.
  • Plate 200 µl of the solution on a proper agar plate.
  • Incubate overnight at 37°C.













  • LB agar plates with proper antibiotic added incubated at 37°C
  • Thawed Invitrogen TOP10 cells (every tube contains approximately 60 µl of competent cells)
  • Resuspended DNA
  • SOC medium