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Team:Paris Liliane Bettencourt/Notebook/2010/08/22/
From 2010.igem.org
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Aleksandra
Gel electrophoresis
- Ps/wRBS/LuxR/attC/t/attC/mRFP
- -
- Ps/mRBS/LuxR/attC/t/attC/mRFP
-
- sRBS/LacZ (wrong enzymes, no interest)
- -
- wRBS/TetA(C)
- -
- mRBS/TetA(C)
- -
- LuxI
- Ladder 1kb
<a href="/index.php?title=Special:Upload&wpDestFile=Gel2208_1.jpg" class="new" title="Image:Gel2208 1.jpg">Image:Gel2208 1.jpg</a>
Gel extraction, elution in 35µl warm water
- LuxI
PCR
1µl miniprep + 2x1µl primers VF2 and VR (0.025ng/µl) + 10µl water + 13µl 2x Quick load mix; Tm=60°C, Telong=72°C (should have been 68°C)
- wRBS/TetA(C)
- mRBS/TetA(C)
- sRBS/TetA(C)
PCR purification kit, elution in 35µl warm water
- wRBS/TetA(C)
- mRBS/TetA(C)
- sRBS/TetA(C)
Digestion with XbaI and SpeI : 15µl PCR product + 2x1µl enzymes + 2µl B3 + 0.2µl BSA, then inactivation 10mins at 80°C
- wRBS/TetA(C)
- mRBS/TetA(C)
- sRBS/TetA(C)
Overnight cultures
- Ps/sRBS/LuxR/attC/t/attC/mRFP
- Ps/mRBS/LuxR/attC/t/attC/mRFP
- Ps/wRBS/LuxR/attC/t/attC/mRFP
