• Insert your content here.

</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>
<img src="/images/c/c3/Resultset_previous.png" border="0" /><a href="/wiki/IGEM:Paris_Liliane_Bettencourt/Notebook/Paris_2010/2010/07/28" title="IGEM:Paris Liliane Bettencourt/Notebook/Paris 2010/2010/07/28">Previous entry</a>      <a href="/wiki/IGEM:Paris_Liliane_Bettencourt/Notebook/Paris_2010/2010/07/30" title="IGEM:Paris Liliane Bettencourt/Notebook/Paris 2010/2010/07/30">Next entry</a><img src="/images/5/5c/Resultset_next.png" border="0" /> </td></tr> <tr>

Raphaël
Making Amp plates
40 plates

Aleksandra and Théotime

Miniprep

• stand.RBS /TetA(C) 1 and 2
• pLux/RBS/GFP/term A and B (transformations from 07/27)
• Pmed/weakRBS/LuxR 1 and 2
• Pstr/weakRBS/LuxR 1 and 2
• Pmed /medRBS/LuxR 1 and 2
• Pstr/medRBS/LuxR 1 and 2
• Pmed/strRBS/LuxR 1 and 2
• Pstr/strRBS/LuxR 1 and 2
• stand.RBS/LacZ 1 and 2
• weak RBS/LacZ 1 and 2
• LuxI/term 1 and 2
• attC
• term/attC/mRFP
• pLux/RBS/GFP/term 1 to 5 (transformations from 07/21)

Théotime and Raphaël
Restriction digest, incubation at 37°C for 2h.
Restriction digests to do gel purification :

• 20µl stand.RBS /TetA(C) 1 + 1µl EcoRI + 1µl SpeI + 2.5µl buffer4 10x + 0.25µl BSA 100x
• 20µl med.RBS /TetA(C) + 1µl EcoRI + 1µl SpeI + 2.5µl buffer4 10x + 0.25µl BSA 100x
• 20µl weak.RBS /TetA(C) + 1µl EcoRI + 1µl SpeI + 2.5µl buffer4 10x + 0.25µl BSA 100x
• 20µl stand.RBS /LacZ 1 + 1µl EcoRI + 1µl SpeI + 2.5µl buffer4 10x + 0.25µl BSA 100x
• 20µl weak.RBS /LacZ 1 + 1µl EcoRI + 1µl SpeI + 2.5µl buffer4 10x + 0.25µl BSA 100x
• 20µl LuxI/term 1 + 1µl EcoRI + 1µl XbaI + 2.5µl buffer4 10x + 0.25µl BSA 100x
• 20µl attC + 1µl SpeI + 1µl PstI + 2.5µl buffer2 10x + 0.25µl BSA 100x
• 20µl term/attC/mRFP + 1µl XbaI + 1µl PstI + 2.5µl buffer3 10x + 0.25µl BSA 100x

Restriction digests to verify the transformations :

• 5µl pLux/GFP 1 + 1µl XbaI + 1µl PstI + 2µl buffer3 10x + 0.2µl BSA 100x + 11µl water
• 5µl pLux/GFP 2 + 1µl XbaI + 1µl PstI + 2µl buffer3 10x + 0.2µl BSA 100x + 11µl water
• 5µl pLux/GFP 3 + 1µl XbaI + 1µl PstI + 2µl buffer3 10x + 0.2µl BSA 100x + 11µl water
• 5µl pLux/GFP 4 + 1µl XbaI + 1µl PstI + 2µl buffer3 10x + 0.2µl BSA 100x + 11µl water
• 5µl pLux/GFP A + 1µl XbaI + 1µl PstI + 2µl buffer3 10x + 0.2µl BSA 100x + 11µl water
• 5µl pLux/GFP B + 1µl XbaI + 1µl PstI + 2µl buffer3 10x + 0.2µl BSA 100x + 11µl water
• 5µl Pstr/weakRBS/LuxR 1 + 1µl XbaI + 1µl PstI + 2µl buffer3 10x + 0.2µl BSA 100x + 11µl water
• 5µl Pmed/weakRBS/LuxR 1 + 1µl XbaI + 1µl PstI + 2µl buffer3 10x + 0.2µl BSA 100x + 11µl water
• 5µl Pstr/medRBS/LuxR 1 + 1µl XbaI + 1µl PstI + 2µl buffer3 10x + 0.2µl BSA 100x + 11µl water
• 5µl Pmed /medRBS/LuxR 1 + 1µl XbaI + 1µl PstI + 2µl buffer3 10x + 0.2µl BSA 100x + 11µl water
• 5µl Pstr/strRBS/LuxR 1 + 1µl XbaI + 1µl PstI + 2µl buffer3 10x + 0.2µl BSA 100x + 11µl water
• 5µl Pmed/strRBS/LuxR 1 + 1µl XbaI + 1µl PstI + 2µl buffer3 10x + 0.2µl BSA 100x + 11µl water

Aleksandra and Raphaël
Gel electrophoresis (1.5% agarose, 100V) Since the yield of the gel purification was bad (between 5 and 30%), the experiment was done at the Necker's lab in order to control the role of materials and buffers.

• pLux/GFP 1 : 20µl + 5µl lb
• pLux/GFP 2 : 20µl + 5µl lb
• pLux/GFP 3 : 20µl + 5µl lb
• pLux/GFP 4 : 20µl + 5µl lb
• pLux/GFP 5 : 20µl + 5µl lb
• pLux/GFP A : 20µl + 5µl lb
• pLux/GFP B : 20µl + 5µl lb
• Pstr/weakRBS/LuxR 1 : 20µl + 5µl lb
• Pmed/weakRBS/LuxR 1 : 20µl + 5µl lb
• Pstr/medRBS/LuxR 1 : 20µl + 5µl lb
• Pmed /medRBS/LuxR 1 : 20µl + 5µl lb
• Ladder 1Kb (Fermentas) : 10 µL

<a href="/wiki/Image:Gel2907_1.jpg" class="image" title="Image:gel2907_1.jpg"><img alt="Image:gel2907_1.jpg" src="/images/e/e4/Gel2907_1.jpg" width="976" height="582" border="0" /></a>

• Pstr/strRBS/LuxR 1 : 20µl + 5µl lb
• Pmed/strRBS/LuxR 1 : 20µl + 5µl lb
• Ladder 1Kb (Fermentas) : 10 µL
• term/attC/mRFP : 25µl + 5µl lb
• -
• weak.RBS /TetA(C) 1 : 25µl + 5µl lb
• -
• med.RBS /TetA(C) 1 : 25µl + 5µl lb
• -
• stand.RBS /TetA(C) 1 : 25µl + 5µl lb
• -
• -

<a href="/wiki/Image:Gel2907_2.jpg" class="image" title="Image:gel2907_2.jpg"><img alt="Image:gel2907_2.jpg" src="/images/8/85/Gel2907_2.jpg" width="917" height="526" border="0" /></a> <a href="/wiki/Image:Gel2907_3.jpg" class="image" title="Image:gel2907_3.jpg"><img alt="Image:gel2907_3.jpg" src="/images/4/49/Gel2907_3.jpg" width="966" height="590" border="0" /></a>

• weak.RBS /LacZ 1 : 25µl + 5µl lb
• -
• stand.RBS /LacZ 1  : 25µl + 5µl lb
• -
• Ladder 100bp : 10 µL
• -

<a href="/wiki/Image:Gel2907_4.jpg" class="image" title="Image:gel2907_4.jpg"><img alt="Image:gel2907_4.jpg" src="/images/e/ee/Gel2907_4.jpg" width="479" height="561" border="0" /></a> <a href="/wiki/Image:Gel2907_5.jpg" class="image" title="Image:gel2907_5.jpg"><img alt="Image:gel2907_5.jpg" src="/images/3/3c/Gel2907_5.jpg" width="463" height="573" border="0" /></a>