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Raphaël
Restriction digest with EcoRI-HF and SpeI : 20µl miniprep + 2x1µl enzyme + 2.5µl B4 + 0.25µl BSA

• Left arm HK022 Mut-Mut
• Left arm HK022 WT-Mut
• Left arm λ Mut-Mut
• Left arm λ WT-Mut
• Left arm WT-WT

Restriction digest with EcoRI-HF and XbaI : 20µl miniprep + 2x1µl enzyme + 2.5µl B4 + 0.25µl BSA

• Kan resistance cassette

Restriction digest with XbaI and PstI : 20µl miniprep + 2x1µl enzyme + 2.5µl B3 + 0.25µl BSA

• Right arm HK022 Mut-Mut
• Right arm HK022 WT-Mut
• Right arm λ Mut-Mut
• Right arm λ WT-Mut

Restriction digest with SpeI and PstI : 20µl miniprep + 2x1µl enzyme + 2.5µl B2 + 0.25µl BSA

• LacZα expression cassette

</td></tr> <tr> <td style="background-color: #EEE"><a href="/wiki/Image:Igem-logo-150px.png" class="image" title="Igem-logo-150px.png"><img alt="" src="/images/7/7b/Igem-logo-150px.png" width="150" height="115" border="0" /></a>Population counter </td><td style="background-color: #F2F2F2" align="center"><img src="/images/9/94/Report.png" border="0" /> <a href="/wiki/IGEM:Paris_Liliane_Bettencourt/Notebook/Paris_2010" title="IGEM:Paris Liliane Bettencourt/Notebook/Paris 2010">Main project page</a>
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Aleksandra
Minipreps

• 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
• pLux
• wRBS/LacZ
• sRBS/LacZ
• wRBS/TetA(C)
• mRBS/TetA(C)
• 2x RBS/LuxI

Raphaël
Restriction digest with XbaI and PstI : 20µl miniprep + 2x1µl enzyme + 2.5µl B2 + 0.25µl BSA

• wRBS/TetA(C)
• mRBS/TetA(C)
• 2x RBS/LuxI

Restriction digest with SpeI and PstI : 20µl miniprep + 2x1µl enzyme + 2.5µl B2 + 0.25µl 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
• pLux
• wRBS/LacZ
• sRBS/LacZ

Aleksandra and Raphaël

Gel electrophoresis

• Ps/wRBS/LuxR + attC/term/attC/mRFP
• Ladder 1kb
• Ps/mRBS/LuxR + attC/term/attC/mRFP
• -
• Ps/sRBS/LuxR + attC/term/attC/mRFP
• -
• Pm/sRBS/LuxR + attC/term/attC/mRFP
• -
• pLux
• -
• wRBS/LacZ

<a href="/index.php?title=Special:Upload&wpDestFile=Gel2008_1.jpg" class="new" title="Image:Gel2008 1.jpg">Image:Gel2008 1.jpg</a>

Gel electrophoresis

• sRBS/LacZ
• -
• wRBS/TetA(C)
• -
• mRBS/TetA(C)
• -
• RBS/LuxI
• RBS/LuxI
• Ladder 1kb
• -
• -
• -

-> The gel was unfortunately damaged... thus we have lost a lot of DNA and the migration was abnormal.

<a href="/index.php?title=Special:Upload&wpDestFile=Gel2008_2.jpg" class="new" title="Image:Gel2008 2.jpg">Image:Gel2008 2.jpg</a>

Léa

Gel extraction

• Ps/sRBS/LuxR + attC/term/attC/mRFP
• Pm/sRBS/LuxR + attC/term/attC/mRFP
• pLux
• wRBS/LacZ