Ligations (V + I)

• 1) pLux + LuxI
• 2) RBS/GFP/tt (E0240) + LuxI
• 3) Pms/mTet/LuxI + I13507/pLux/GFP
• 4) Ctrl pLux
• 5) Ctrl RBS/GFP/tt (E0240)
• 6) Ctrl Pms/mTet/LuxI

Transformation of 1-6 + positive control

Minipreps

• PmS-MTet-LuxI
• WTet
• MTet
• STet
• Ps
• pSB1C3 PmS-MTet-LuxI I
• pSB1C3 PsS-WTet-LuxI1 I
• PsS-WTet-LuxI2
• PsS-WTet-LuxI1
• pSB1C3 PsS-WTet-LuxI2 I
• pSB1C3 PmS-MTet-LuxI II

Restriction digest

With XbaI and PstI

• WTet
• MTet
• STet

With SpeI and PstI

• Ps

With EcoRI-HF and SpeI

• pSB1C3 PmS-MTet-LuxI I
• pSB1C3 PsS-WTet-LuxI1 I
• pSB1C3 PsS-WTet-LuxI2 I
• pSB1C3 PmS-MTet-LuxI II
• PmS-MTet-LuxI

Gel electrophoresis

• WTet
• MTet
• STet
• pSB1C3 PmS-MTet-LuxI I
• pSB1C3 PsS-WTet-LuxI1 I
• pSB1C3 PsS-WTet-LuxI2 I
• pSB1C3 PmS-MTet-LuxI II
• PmS-MTet-LuxI

Gel extraction

• WTet
• MTet
• STet

PCR purification

• Ps

Transformation

• PsS-WTet + pBad/Int
• PmS-MTet + pBad/Int
• positive control

Overnight cultures

• PmS-MTet-LuxI
• PsS-WTet-LuxI
• PmS Chr
• LuxI
• attP
• PsS-RFP
• PsW-RFP
• PsM-RFP
• PmS-RFP
• pBad/Int

From transformations of this morning :

• Final PmS-MTet x5
• LuxI/GFP x1
• pLux/LuxI x2 -> one colony didnt' grow