Team:Marburg/Notebook:March

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Revision as of 12:19, 14 August 2013

Notebook


<a name="11-11-2012">11.11.2011</a>

<fieldset class="experiment general"> 

   <legend><a name="title">Überschrift</a></legend>

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.

</fieldset>


<a name="17-04-2013">17.04.2013</a>

<fieldset class="experiment ligation"> 

   <legend><a name="lig">Ligation</a></legend>

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

</fieldset>

<fieldset class="experiment digest"> 

   <legend><a name="dig">Digest</a></legend>

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

<colgroup> <col width="50%" /> <col width="50%" /> </colgroup> <thead> </thead> <tbody> </tbody>
Gel electrophoresis

<img src="beispiel.png" width="50%" alt="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.

</fieldset>


<a name="18-04-2012">18.04.2012</a>

<fieldset class="experiment transformation"> 

   <legend>Transformation</legend>

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 .

</fieldset> <fieldset class="experiment pcr"> 

   <legend>PCR</legend>

Investigator: Patrick, Lucas

Aim: Receive PvuII point-mutation on colonies 3 and 5 of E. coli pB201-40JO.

<colgroup> <col width="15%" /> <col width="30%" /> <col width="5%" /> <col width="20%" /> <col width="20%" /> <col width="10%" /> </colgroup> <thead>

</thead>

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


<colgroup> <col width="50%" /> <col width="50%" /> </colgroup> <thead> </thead> <tbody> </tbody>
Gel electrophoresis

<img src="beispiel.png" width="50%" alt="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.

</fieldset>


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