Team:Marburg/Notebook:March
From 2013.igem.org
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
</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
Gel electrophoresis | |
---|---|
<img src="beispiel.png" width="50%" alt="gel-electrophoresis-image" /> |
Gel substances
Expactations
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.
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 | |
---|---|
<img src="beispiel.png" width="50%" alt="gel-electrophoresis-image" /> |
Gel substances
Expactations
We don’t receive all expected fragments. The expected fragment in lane 1 is missing. |
</fieldset>