Team:Goettingen/NoteBook

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===Timeline===
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===Notebook===
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This year we started early. At the beginning of January 2013, 12 of us gathered together and formed our [[Team:Goettingen/Team/Organization|Organization Team]]. With all these brain storming and background research. We decided to look into the topic "Antibiotic Resistance", which is less known to the public, but actually very urgent problem. We designed our team logo and finished the booklet for fund-raising ([[Team:Goettingen/Outreach|more documents please visit our Outreach page]]). Finally, we determined the title of our project, "The beast and its Achilles heel: a novel target to fight multi-resistant pathogenic bacteria".
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<div class="monat">June</div>
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Shortly before May, our lab work started and we have had six more students on the team. We divided our team into 3 subteams, each team has carried out one dimension of our project. After four full month of hard work, we finally wrapped up, and proudly present you all the results on this team wiki.
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<div class="tlob" id="tl_0706">
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Click the icons on the right side to check out our timeline or our standard protocol.
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<span class="date">07th<img src="https://static.igem.org/mediawiki/2013/b/b7/Goe-timeline-dot.png" /></span>
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<div class="cont">
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<p class="timeline-title">Plasmid mini-prep for Part1-7</p>
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<b>Cryo-Stocks of E. coli transformants (C1, parts 1 – 7):</b>
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<ul>
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<li>900 μl E.coli ON culture + 100 μl DMSO 100% in special tubes (ask Katrin or Katrin^^)</li>
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<li>vortex</li>
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<li>store at – 70 °C (red box)</li>
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</ul>
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<b>Plasmid Mini-Preparation of parts 1 - 7:</b>
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<p>ON cultures of C1 for parts 1 – 7 with kit “NucleoSpin? Plasmid” von Macherery-Nagel according to manual pp. 16-17 (NucleoSpin? Plasmid protocol for purification of high copy plasmids</p>
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<p>Step 5: recommended washing of silica membrane with buffer AW was performed</p>
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<p>Step 7: Elution with buffer AE pre-heated to 50 – 60 °C (in future: DON’T elute with this buffer, use pre-heated HPLC-H2O instead! No one knows what’s inside the buffer and its components (EDTA?) could interfere with sequencing and other reactions)</p>
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<br / >
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<p>NanoDrop – Plasmid concentrations</p>
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<table cellspacing="0">
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<tr><th>Part Number</th><th>c(DNA)[ng/μl]</th><th>A<sub>260</sub>/A<sub>280</sub></th><th>A<sub>260</sub>/A<sub>230</sub></th>
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<tr><td>1</td><td>84.6</td><td>1.94</td><td>2.18</td></tr>
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<tr><td>2</td><td>79.8</td><td>1.94</td><td>2.10</td></tr>
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<tr><td>3</td><td>151.0</td><td>1.89</td><td>2.24</td></tr>
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<tr><td>4</td><td>29.5</td><td>1.91</td><td>2.07</td></tr>
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<tr><td>5</td><td>154.9</td><td>1.88</td><td>2.25</td></tr>
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<tr><td>6</td><td>88.9</td><td>1.92</td><td>2.08</td></tr>
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<tr><td>7</td><td>5.9</td><td>14.08</td><td>1.87</td></tr>
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<p>Stored in red box at - 20°C</p>
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<b>Primers arrived!</b>
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<p>iGEM_32 ~ 37: dissolved in HPLC water, stored at -20oC in our box</p>
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<p>100uM stock , for PCR dilute 1:20 in HPLC water.</p>
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<p>Primer 32 and 33 are strong in forming 2nd structures, increase the amount in PCR.</p>
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<a href="/Team:Goettingen/NoteBook/timeline" style="margin-left:15px;display:inline-block;text-decoration:none;">
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<h2 style="text-align:center">Timeline</h2>
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<img src="https://static.igem.org/mediawiki/2013/a/ae/Goe-timeline-icon.png">
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<div class="tlob" id="tl_0606">
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<a href="https://static.igem.org/mediawiki/2013/e/eb/Goe-iGEM_Protocolls.pdf" style="margin-left:50px;display:inline-block;text-decoration:none;">
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<span class="date">06th<img src="https://static.igem.org/mediawiki/2013/b/b7/Goe-timeline-dot.png" /></span>
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<h2 style="text-align:center">Protocol</h2>
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<div class="cont">
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<img src="https://static.igem.org/mediawiki/2013/3/32/Goe-protocol.png">
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<p class="timeline-title">Pick the colonies of part1-7</p>
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</a>
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<b>Media preparation</b>
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<p>1000ml LB+Amp Solid medium => about 50 Plates(with black code)</p>
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<b>Transformation</b>
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<tr><th>Entry Numer</th><th>Location on the kit</th></tr>
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<tr><td>BBa_B0034</td><td>P5 2 M</td></tr>
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<b>Pick the colonies of Part1-7</b>
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<p>4ml LB with antibiotics, overnight culture for mini-prep[marked with C1]</p>
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<p>Backup plates:marked with C1,C2,C3 for each part.</p>
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<br><br>
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<span class="date">05th<img src="https://static.igem.org/mediawiki/2013/b/b7/Goe-timeline-dot.png" /></span>
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<div class="cont">
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<p class="timeline-title">Preparation of the medium, antibioticks, Transformation.</p>
 
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<b>Preparation of Antibiotic Stocks</b>
 
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<ul>
 
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<li>1000x Ampicillin 10 1mL Stocks (EPs in the red box in -20 freezer)</li>
 
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<li>1000x Chloramphenicol 10mL Stock (Falcon in Freezer)</li>
 
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</ul>
 
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<b>Media preparation</b>
 
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<ul><li>250ml*4 LB media with Cm </li>
 
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<li>250ml*1 LB media with Amp </li>
 
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<b>Primer design</b>
 
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<table cellspacing="0">
 
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<tr><th>Code Name</th><th>  Description</th></tr>
 
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<tr><td>iGEM_36</td><td>  DarR operator sequence + biobrick prefix</td></tr>
 
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<tr><td>iGEM_37</td><td>  DarR operator sequence + biobrick sufix</td></tr>
 
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</table><br />
 
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<b>Transformation</b>
 
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<tr><th>Biobrick entry number</th><th>Mark</th></tr>
 
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<tr><td>BBa_J23117</td><td>  part 1</td></tr>
 
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<tr><td>BBa_J23116</td><td>  part 2</td></tr>
 
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<tr><td>BBa_J23110</td><td>  part 3</td></tr>
 
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<tr><td>BBa_J23118</td><td>  part 4</td></tr>
 
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<tr><td>BBa_J61101</td><td>  part 5</td></tr>
 
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<tr><td>BBa_BE0240-Cm</td><td>  part 6 -- Cm</td></tr>
 
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<tr><td>BBa_B0015</td><td>  part 7 -- Cm</td></tr><tr>
 
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<td>BBa_B0034</td><td>  part 8</td></tr>
 
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</table>
 
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Resuspended but not transformed
 
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<table>
 
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<tr><th>Entry number</th><th>  location on kit</th></tr>
 
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<tr><td>BBa_E0204-Amp</td><td>  P5 12 M</td></tr>
 
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<tr><td>BBa_QO3121</td><td>  P5 20 N</td></tr>
 
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</table>
 
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</div>
 
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<div class="tlob" id="tl_0406">
 
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<span class="date">04th<img src="https://static.igem.org/mediawiki/2013/b/b7/Goe-timeline-dot.png" /></span>
 
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<div class="cont">
 
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<p class="timeline-title">Find the correct DNA sequence of DarR and primer design</p>
 
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<div class="timeline-cont">
 
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<table cellspacing="0">
 
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<tr><th>Code Name</th><th>  Description</th></tr>
 
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<tr><td>iGEM_32</td><td>  Forward primer for DarR ORF amplification, with biobrick prefix</td></tr>
 
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<tr><td>iGEM_33</td><td>  Reverse primer for DarR ORF amplification, with biobrick sufix</td></tr>
 
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<tr><td>iGEM_34</td><td>  Forward primer for DarR ORF PCR amplification sequencing.</td></tr>
 
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<tr><td>iGEM_35</td><td>  Reverse primer for DarR ORF PCR amplification sequencing.</td></tr>
 
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</table>
 
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Latest revision as of 19:24, 3 October 2013





The beast and its Achilles heel:

 A novel target to fight multi-resistant pathogenic bacteria


Notebook

This year we started early. At the beginning of January 2013, 12 of us gathered together and formed our Organization Team. With all these brain storming and background research. We decided to look into the topic "Antibiotic Resistance", which is less known to the public, but actually very urgent problem. We designed our team logo and finished the booklet for fund-raising (more documents please visit our Outreach page). Finally, we determined the title of our project, "The beast and its Achilles heel: a novel target to fight multi-resistant pathogenic bacteria".

Shortly before May, our lab work started and we have had six more students on the team. We divided our team into 3 subteams, each team has carried out one dimension of our project. After four full month of hard work, we finally wrapped up, and proudly present you all the results on this team wiki.

Click the icons on the right side to check out our timeline or our standard protocol.

Timeline

Protocol

Retrieved from "http://2013.igem.org/Team:Goettingen/NoteBook"