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Aleksandra
Gel extraction

• Ps/sRBS/LuxR/attC/term/attC/mRFP
• Pm/sRBS/LuxR/attC/term/attC/mRFP
• I13507/pLux/GFP
• wRBS/tetA(C)
• mRBS/tetA(C)
• sRBS/tetA(C)
• sRBS/LacZα/LuxI III (from )
• pLux/LuxI II
• Ps/mRBS/LuxR/attC/term/attC/wRBS/TetA(C) II
• Ps/mRBS/LuxR/attC/term/attC/wRBS/TetA(C) III

Raphaël
Minipreps

• attP-E (pSWK)
• Pm/sRBS/LuxR/attC/term/attC/mRFP

Restriction digest
With EcoRI-HF and PstI (15µl DNA + 2µl B2 + 2x1µl enzymes + 0.2µl BSA):

• attP-E (pSWK)

With EcoRI-HF and PstI and AlWNI (10µl DNA + 5µl water + 2µl B2 + 3x1 µl enzyme + 0.2µl BSA)

• Ps/wRBS/LuxR/attC/term/attC/mRFP
• Ps/mRBS/LuxR/attC/term/attC/mRFP
• Ps/sRBS/LuxR/attC/term/attC/mRFP
• Pm/sRBS/LuxR/attC/term/attC/mRFP

Gel electrophoresis

• attP-E (pSWK)
• -
• Ps/wRBS/LuxR/attC/term/attC/mRFP
• -
• Ps/mRBS/LuxR/attC/term/attC/mRFP
• -
• Ps/sRBS/LuxR/attC/term/attC/mRFP
• -
• Pm/sRBS/LuxR/attC/term/attC/mRFP
• -
• Ladder 1 kb
• -
• -
• -
• -
• -
• -

<a href="/index.php?title=Special:Upload&wpDestFile=Gel0209_1.jpg" class="new" title="Image:Gel0209 1.jpg">Image:Gel0209 1.jpg</a>

Gel extraction

• attP-E (pSWK)
• Ps/mRBS/LuxR/attC/term/attC/mRFP
• Ps/sRBS/LuxR/attC/term/attC/mRFP
• Pm/sRBS/LuxR/attC/term/attC/mRFP

Ligations (V + I)

• 1) attP + Pms
• 2) attP + Psm
• 3) attP + Pss
• 4) pLux + LuxI
• 5) wlacZα + LuxI
• 6) sLacZα + LuxI
• 7) Psm + slacZα/LuxI III
• 8) Psm + wTet/LuxI
• 9) Pss + wTet/LuxI
• 10) Pms + wTet/LuxI
• 11) I13507/pLux/GFP + Psm/wTet II
• 12) I13507/pLux/GFP + Psm/wTet III
• 13) mTet/LuxI + Psm
• 14) sTet/LuxI + Psm
• 15) sLacZα/LuxI III + Pss
• 16) mTet/LuxI + Pss
• 17) sTet/LuxI + Pss
• 18) sLacZα/LuxI III + Pms
• 19) mTet/LuxI + Pms
• 20) sTet/LuxI + Pms
• A) ctrl attP
• B) ctrl pLux
• C) ctrl wLacZα
• D) ctrl sLacZα
• E) ctrl Psm
• F) ctrl Pss
• G) ctrl Pms
• H) ctrl I13507

Aleklsandra
Transformation of 1-20, A-H, plating with 150 to 200µl

Overnight cultures

• Ps/wRBS/LuxR/attC/term/attC/mRFP and pBAD/Int (double transformants)
• Ps/mRBS/LuxR/attC/term/attC/mRFP and pBAD/Int (double transformants)
• Ps/sRBS/LuxR/attC/term/attC/mRFP and pBAD/Int (double transformants)
• wRBS/tetA(C)
• mRBS/tetA(C)
• sRBS/tetA(C)
• wRBS/LacZα
• sRBS/LacZα
• Ps