Team:Marburg/Notebook:Template

From 2013.igem.org

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<a name="11-11-2012">11.11.2011</a>
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<a name="11-11-2012">11.11.2011</a>
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    <legend><a name="title">Überschrift</a></legend>
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<span class="inv-names">X,Y</span>
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<span class="aim-desc">We attempt to get results.</span>
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<span class="aim-desc">We attempt to get results.</span>
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<p>Wichtig ist, dass immer die verwendete Materialien (Plasmide, genomische DNA [aus welchen Stamm], Primer) genau benannt und/oder ggf. irgendwo nummeriert werden. Es sollte alles nachvollziehbar sein, wann ihr mit welchen Proben was gemacht habt.</p>
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<p>Wichtig ist, dass immer die verwendete Materialien (Plasmide, genomische DNA [aus welchen Stamm],  
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<p>Auf den folgenden Seiten sind Beispiele, wie wir uns das Notebook vom Aufbau und Detailgrad vorstellen.</p>
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Primer) genau benannt und/oder ggf. irgendwo nummeriert werden. Es sollte alles nachvollziehbar sein,  
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wann ihr mit welchen Proben was gemacht habt.</p>
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<p>Auf den folgenden Seiten sind Beispiele, wie wir uns das Notebook vom Aufbau und Detailgrad vorstellen.</p>
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<li>5 µl vector DNA (pSB1C3)</li>
<li>5 µl vector DNA (pSB1C3)</li>
<li>20 µl insert DNA (1, 3, 4, 5, 6, 7 or 8)</li>
<li>20 µl insert DNA (1, 3, 4, 5, 6, 7 or 8)</li>
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<li>4 µl DNA (pSB1C3-J04450)</li>
<li>4 µl DNA (pSB1C3-J04450)</li>
<li>1 µl MluI</li>
<li>1 µl MluI</li>
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<p>We don’t receive all expected fragments. The expected fragment in lane 1 is missing.</p>
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<p>We don’t receive all expected fragments. The expected fragment in lane 1 is missing.</p>
<p>We don’t receive all expected fragments. The expected fragment in lane 1 is missing.</p>
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Revision as of 17:10, 26 September 2013

Notebook

11.11.2011

Überschrift
Investigator: X,Y
Aim: We attempt to get results.

Wichtig ist, dass immer die verwendete Materialien (Plasmide, genomische DNA [aus welchen Stamm], Primer) genau benannt und/oder ggf. irgendwo nummeriert werden. Es sollte alles nachvollziehbar sein, wann ihr mit welchen Proben was gemacht habt.

Auf den folgenden Seiten sind Beispiele, wie wir uns das Notebook vom Aufbau und Detailgrad vorstellen.

17.04.2013

Ligation
Investigator: Franzi, Lucas
Aim: Assemble the Bricks 1, 3, 4, 5, 6, 7 and 8 into the vector pSB1C3.
  • 5 µl vector DNA (pSB1C3)
  • 20 µl insert DNA (1, 3, 4, 5, 6, 7 or 8)
  • 3 µl 10x T4 DNA ligase buffer
  • 2 µl T4 DNA ligase

The samples were incubated for 14 h at 18° C.

Digest
Investigator: Christian, Patrick
Aim: Digest of pSB1C3-J04450 with MluI and HindIII.
  • 4 µl DNA (pSB1C3-J04450)
  • 1 µl MluI
  • 1 µl HindIII
  • 1,5 µl 10x red buffer
  • 1,5 µl 10x orange g loading buffer
  • 6 µl H2O

The samples were incubated for 4 h at 37° C.

Gel electrophoresis
gel-electrophoresis-image

Gel substances

  • 1% Agarose gel
  • 10 µl RedSafe in 50 ml gel
  • x µl Hyper Ladder

Expactations

  • Lane 1: 5300 kbp
  • Lane 2: 3300 kbp
  • Lane 3: 1300 kbp

We don’t receive all expected fragments. The expected fragment in lane 1 is missing.

We don’t receive all expected fragments. The expected fragment in lane 1 is missing.

18.04.2012

Transformation
Investigator: Franzi, Christian
Aim: Transformation of the plasmid DNA pB201-40JO in E. coli for amplification.

The chemo competent E. coli DH5a cells were transformed with pB201-40JO DNA and plated on dYT-Amp-plates.
The plates were incubated over night at 37° C .

PCR
Investigator: Patrick, Lucas
Aim: Receive PvuII point-mutation on colonies 3 and 5 of E. coli pB201-40JO.
Volume Reagent   Temp (°C) Time
10 µl 5x Buffer   95 3 min
1,5 µl Primer fwd 13   95 3 sec
1,5 µl Primer rev 14   58 30 sec x17
1 µl Template   72 1 min
36 µl H2O   7 min 3 min
1 µl Phusion Phusion Polymerase   4 Hold

Gel electrophoresis
gel-electrophoresis-image

Gel substances

  • 1% Agarose gel
  • 10 µl RedSafe in 50 ml gel
  • x µl Hyper Ladder

Expactations

  • Lane 1: 5300 kbp
  • Lane 2: 3300 kbp
  • Lane 3: 1300 kbp

We don’t receive all expected fragments. The expected fragment in lane 1 is missing.

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