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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)