Team:Paris Liliane Bettencourt/Notebook/2010/09/02/
From 2010.igem.org
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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