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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