IGEM:Paris Liliane Bettencourt/Notebook/Paris 2010/2010/07/16

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Stéphane
Gel electrophoresis (agarose 1,0% w/v, EtB) Verification of the DNA quantity at each step from the last batch (minipreps, restriction and ligation of 07/15).

  • 6 µL DNA ladder (C#83 µg/mL)
  • 5 µL pLux (R0062) digest with SpeI and PstI + 1 µL loading buffer
  • nothing
  • 5 µL pLux (R0062) after gel extraction + 1 µL loading buffer
  • 5 µL RBS/GFP/term (E0240) digest with XbaI and PstI + 1 µL loading buffer
  • nothing
  • 5 µL RBS/GFP/term (E0240) after gel extraction + 1 µL loading buffer
  • 5 µL pLux - RBS/GFP/term ligation + 1 µL loading buffer

Image:verificationgel.jpg



Overnight culture
In 10ml LB and 10 µL corresponding antibiotic

  • attC - Kan
  • term-attC-mRFP - Kan
  • term (B0015) - Amp
  • RBS/GFP/term (E0240) - Amp
  • RBS/LuxR (J37033) - Amp
  • pLux (R0062) - Amp
  • RBS/LuxI (K081008) - Amp
  • Pstrong (J23119) - Amp
  • Pmedium (J23110) - Amp
  • kanR (P1003) - Amp


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