Team:UNIPV-Pavia/Methods Materials/Transformation

From 2009.igem.org

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*'''LB agar plates with proper antibiotic'''
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*'''Invitrogen TOP10 cells'''
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'''LB agar plates with proper antibiotic'''<hr color='white' width='70%'>
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*'''Resuspended DNA'''
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'''Invitrogen TOP10 cells'''<hr color='white' width='70%'>
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*'''SOC medium'''
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'''Resuspended DNA'''<hr color='white' width='70%'>
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'''SOC medium'''</font>
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Revision as of 16:26, 22 September 2009

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 thaw TOP10 cells tube (every tube contains approximately 60 µl of competent cells).
  • 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
Invitrogen TOP10 cells
Resuspended DNA

SOC medium





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