Team:Marburg/Notebook:July

From 2013.igem.org

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<div class="notebooky-entry">
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<h2 class="title">
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<a name="02-07-2013">02.07.2013</a>
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</h2>
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<fieldset class="experiment miniprep">
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    <legend><a name="min">Miniprep</a></legend>
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    <div class="investigator">
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<span class="inv">Investigator:</span>
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<span class="inv-names">Dominik</span>
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</div>
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    <div class="aim">
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<span class="aim">Aim:</span>
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<span class="aim-desc">preparation of the plasmids pSB1A3-iBB10+iBB9, pSB1A3-iBB11+iBB9, pSB1A3-iBB12+iBB9 and pSB1A3-iBB13+iBB9.</span>
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</div>
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<div class="exp-content">
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<p>The plasmids were isolated with the “QIAprep Spin Miniprep Kit” (Qiagen, Düsseldorf) according to the producer instructions.</p>
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</div>
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</fieldset>
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<fieldset class="experiment digest">
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    <legend><a name="dig">Digest</a></legend>
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    <div class="investigator">
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<span class="inv">Investigator:</span>
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<span class="inv-names">Dominik</span>
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</div>
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    <div class="aim">
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<span class="aim">Aim:</span>
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<span class="aim-desc">Test digest of the plasmids pSB1A3-iBB10+iBB9, pSB1A3-iBB11+iBB9, pSB1A3-iBB12+iBB9 and pSB1A3-iBB13+iBB9 with <i>Eco</i>RI and <i>Pst</i>I</span>
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</div>
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<div class="exp-content">
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<p><ul class="digest">
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<li>1 µl Plasmid (pSB1A3-iBB10+iBB9, pSB1A3-iBB11+iBB9, pSB1A3-iBB12+iBB9 and pSB1A3-iBB13+iBB9)</li>
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<li>0.3 µl EcoRI</li>
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<li>0.3 µl PstI</li>
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<li>1 µl CutSmart</li>
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<li>7.4 µl ddH2O</li>
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</ul>
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<p>The samples were incubated for 1 h at 37° C.</p>
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<table class="gel digest">
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<colgroup>
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<col width="50%" />
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<col width="50%" />
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</colgroup>
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<thead>
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<tr>
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<th colspan="2" class="title">Gel electrophoresis</th>
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</tr>
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</thead>
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<tbody>
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<tr>
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<td>
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<img src="beispiel.png" width="50%" alt="gel-electrophoresis-image" />
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</td>
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<td>
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<p>
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<span class="gel-elc">Gel substances</span>
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<ul class="gel-sub">
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<li>1% Agarose gel</li>
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<li>10 µl RedSafe in 50 ml gel</li>
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<li>x µl Hyper Ladder</li>
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</ul>
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</p>
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<p>
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<span class="exp">Expactations</span>
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<ul class="exp">
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<li>Lane 1: 5300 kbp</li>
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<li>Lane 2: 3300 kbp</li>
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<li>Lane 3: 1300 kbp</li>
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</ul>
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</p>
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<p>Worked as expected.</p>
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</td>
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</tr>
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</tbody>
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</table>
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</div>
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</fieldset>
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<fieldset class="experiment sequencing">
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    <legend>Sequencing</legend>
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    <div class="investigator">
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<span class="inv">Investigator:</span>
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<span class="inv-names">Dominik</span>
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</div>
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    <div class="aim">
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<span class="aim">Aim:</span>
 +
<span class="aim-desc">Analysis of the sequence of pSB1A3-iBB496315.</span>
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</div>
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<div class="exp-content">
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<p>Obviously the wrong plasmid was sent out for sequencing. Due to the lack of enough plasmids for an additional sequence analysis, we will redo a new miniprep.</p>
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</div>
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</fieldset>
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<fieldset class="experiment transformation">
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    <legend>Transformation</legend>
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    <div class="investigator">
 +
<span class="inv">Investigator:</span>
 +
<span class="inv-names">Dominik</span>
 +
</div>
 +
    <div class="aim">
 +
<span class="aim">Aim:</span>
 +
<span class="aim-desc">Transformation of <i>E. coli</i> DH5&#945; with the plasmid DNA pSB1A3-iBB4+iBB10+iBB96315 and pSB1A3-iBB4+iBB13+iBB96315.</span>
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</div>
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<div class="exp-content">
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<p>The chemo competent <i>E. coli</i> DH5&alpha; cells were transformed with the plasmids pSB1A3-iBB4+iBB10+iBB96315 and pSB1A3-iBB4+iBB13+iBB96315 and plated on LB-Amp-plates.<br />
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The plates were incubated over night at 37° C.</p>
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</div>
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</fieldset>
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</div>
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</html>
</html>
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Revision as of 22:09, 30 September 2013

Notebook: July

02.07.2013

Miniprep
Investigator: Dominik
Aim: preparation of the plasmids pSB1A3-iBB10+iBB9, pSB1A3-iBB11+iBB9, pSB1A3-iBB12+iBB9 and pSB1A3-iBB13+iBB9.

The plasmids were isolated with the “QIAprep Spin Miniprep Kit” (Qiagen, Düsseldorf) according to the producer instructions.

Digest
Investigator: Dominik
Aim: Test digest of the plasmids pSB1A3-iBB10+iBB9, pSB1A3-iBB11+iBB9, pSB1A3-iBB12+iBB9 and pSB1A3-iBB13+iBB9 with EcoRI and PstI

  • 1 µl Plasmid (pSB1A3-iBB10+iBB9, pSB1A3-iBB11+iBB9, pSB1A3-iBB12+iBB9 and pSB1A3-iBB13+iBB9)
  • 0.3 µl EcoRI
  • 0.3 µl PstI
  • 1 µl CutSmart
  • 7.4 µl ddH2O

The samples were incubated for 1 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

Worked as expected.
Sequencing
Investigator: Dominik
Aim: Analysis of the sequence of pSB1A3-iBB496315.

Obviously the wrong plasmid was sent out for sequencing. Due to the lack of enough plasmids for an additional sequence analysis, we will redo a new miniprep.

Transformation
Investigator: Dominik
Aim: Transformation of E. coli DH5α with the plasmid DNA pSB1A3-iBB4+iBB10+iBB96315 and pSB1A3-iBB4+iBB13+iBB96315.

The chemo competent E. coli DH5α cells were transformed with the plasmids pSB1A3-iBB4+iBB10+iBB96315 and pSB1A3-iBB4+iBB13+iBB96315 and plated on LB-Amp-plates.
The plates were incubated over night at 37° C.

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