IGEM:Paris Liliane Bettencourt/Notebook/Paris 2010/2010/08/26

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Aleksandra and Raphaël Transformations of Turbocells PmS-WTet PmS-MTet PsS-MTet WLac-LuxI SLac-LuxI pLux-LuxI Controls : pLux SLac WLac PmS PsS pUC19 (positive control) -> almost no colonies on the plates (problem with Turbocells) Colony PCR Touchdown PCR Primers : VF2 + VR WLac-LuxI PsM-MTet I PsM-STet IV PsS-MTet I PsM-WTet I PsM-MTet III PsS-STet IV PsS-WTet III PsM-WTet II PsM-STet II PsS-MTet IV PsS-WTet IV PmS-STet I PmS-WTet I PmS-STet III pLux-GFP (positive control) Minipreps WLac-LuxI PsM-MTet I PsM-STet IV PsS-MTet I PsM-WTet I PsM-MTet III PsS-STet IV PsS-WTet III PsM-WTet II PsM-STet II PsS-MTet IV PsS-WTet IV PmS-STet I PmS-WTet I PmS-STet III Restriction digest with SpeI and EcoRI-HF : 20µl miniprep + 2.5µl B4 + 2x1µl enzymes + 0.25µl BSA WLac-LuxI PsM-MTet I PsM-STet IV PsS-MTet I PsM-WTet I PsM-MTet III PsS-STet IV PsS-WTet III PsM-WTet II PsM-STet II PsS-MTet IV PsS-WTet IV PmS-STet I PmS-WTet I PmS-STet III Restriction digest with XbaI and EcoRI-HF : 20µl miniprep + 2.5µl B4 + 2x1µl enzymes + 0.25µl BSA I13507-pLux-GFP Gel electrophoresis (Colony PCR) WLac-LuxI PsM-MTet I PsM-STet IV PsS-MTet I PsM-WTet I PsM-MTet III PsS-STet IV PsS-WTet III PsM-WTet II PsM-STet II PsS-MTet IV PsS-WTet IV PmS-STet I PmS-WTet I PmS-STet III pLux-GFP Ladder 1Kb Gel electrophoresis (Restriction digest) PmS-STet III Ladder 1Kb PsM-MTet II Ladder 1Kb PsM-STet II Ladder 1Kb WLac-LuxI - PmS-STet I - PsM-MTet I - Gel electrophoresis (Restriction digest) PsS-WTet III Ladder 1Kb PsS-MTet I - PsS-MTet IV - PsS-WTet IV - PsM-WTet II - Gel electrophoresis (Restriction digest) PmS-WTet I Ladder 1Kb PsM-STet IV - PsM-WTet I - I13507-pLux-GFP -> Electrophoresis boxes were not functioning normally and migration could not be completed Transformations of DH5α (because Turbocells didn't work) PmS-WTet PmS-MTet PsS-MTet WLac-LuxI SLac-LuxI pLux-LuxI PsM-MTet PsM-WTet PsW-MTet PsW-WTet PsS-WTet Controls : pLux SLac WLac PmS PsS PsM PsW pUC19 (positive control)