Team:Goettingen/week16-3

From 2012.igem.org

(Difference between revisions)
Line 38: Line 38:
<td width="900" bordercolor="black" valign="top">
<td width="900" bordercolor="black" valign="top">
<h2><b>V08_14 </b></h2><br>
<h2><b>V08_14 </b></h2><br>
-
<b>V08_14 2<sup>nd</sup> round: PCR purification</b><br>
+
<b>2<sup>nd</sup> round: PCR purification</b><br>
<ul>
<ul>
<li>Experiment: <br>The PCR purification was performed using the peqGOLD Cycle-pure Kit (PeqLab), modified using columns from the peqGOLD Plasmid Miniprep Kit (PeqLab)! Elution in 100 µL EB.</li>
<li>Experiment: <br>The PCR purification was performed using the peqGOLD Cycle-pure Kit (PeqLab), modified using columns from the peqGOLD Plasmid Miniprep Kit (PeqLab)! Elution in 100 µL EB.</li>
Line 52: Line 52:
<td width="900" bordercolor="black" valign="top">
<td width="900" bordercolor="black" valign="top">
<h2><b>V08_15 </b></h2><br>
<h2><b>V08_15 </b></h2><br>
-
<b>V08_15 2<sup>nd</sup> round: PCR purification</b><br>
+
<b>V08_15_1 2<sup>nd</sup> round: <i>Dpn</i>I/<i>Bsa</i>I digest and purification</b><br>
 +
<ul>
 +
<li>Experiment: <br>The digest was performed according to protocol from <a href="https://2012.igem.org/Team:Goettingen/week10-3">week 10</a>. The subsequent purification was carried out using the peqGOLD Cycle-pure Kit (PeqLab)!!!</li>
 +
</ul>
 +
<ul>
 +
<li>Observations & Results: <br>The corresponding agarose gel showed band of the expected sizes! At last, there did not seem to be any loss of DNA material.</li>
 +
</ul>
 +
<br>
 +
<b>V08_15_2 2<sup>nd</sup> round: Ligation, 2000 µL</b><br>
 +
<ul>
 +
<li>Experiment: <br>The ligation was performed according to the establoshed protocol from <a href="https://2012.igem.org/Team:Goettingen/week10-3">week 10</a>. Incubation over night at 16 °C.</li>
 +
</ul>
 +
<br></td></tr>
 +
</table>
 +
<br>
 +
<table cellpadding="20 px" border="1" bordercolor="black" valign="top">
 +
<tr bordercolor="black" valign="top">
 +
<td width="900" bordercolor="black" valign="top">
 +
<h2><b>V08_16 </b></h2><br>
 +
<b>V08_16_1 2<sup>nd</sup> round: Purification of ligation from V08_15</b><br>
 +
<ul>
 +
<li>Experiment: <br>The purification of the ligation was now done using the peqGOLD Cycle-pure Kit (PeqLab) instead of an ethanol precipitation. </li>
 +
</ul>
 +
<ul>
 +
<li>Observations & Results: <br>We now seemed to have found the mistake that caused problems with the loss of DNA material in the second mutagenesis round! Miscommunication led to using the wrong kit for purification steps. We decided to use the peqGOLD Cycle-pure Kit (PeqLab) instead of the subsequent ethanol precipitation as well, as this seemed to rise fewer problems.</li>
 +
</ul>
 +
<br>
 +
<b>V08_16_2 2<sup>nd</sup> round: Ligation</b><br>
 +
<ul>
 +
<li>Experiment: <br>The PCR purification was performed using the peqGOLD Cycle-pure Kit (PeqLab), modified using columns from the peqGOLD Plasmid Miniprep Kit (PeqLab)! Elution in 100 µL EB.</li>
 +
</ul>
 +
<ul>
 +
<li>Observations & Results: <br>We now seemed to have found the mistake that caused problems with the loss of DNA material in the second mutagenesis round! Miscommunication led to using the wrong kit for purification steps. We decided to use the peqGOLD Cycle-pure Kit (PeqLab) instead of the subsequent ethanol precipitation as well, as this seemed to rise fewer problems.</li>
 +
</ul>
 +
<br></td></tr>
 +
</table>
 +
<br>
 +
<table cellpadding="20 px" border="1" bordercolor="black" valign="top">
 +
<tr bordercolor="black" valign="top">
 +
<td width="900" bordercolor="black" valign="top">
 +
<h2><b>V08_17 </b></h2><br>
 +
<b>V08_17_1 2<sup>nd</sup> round: <i>Dpn</i>I/<i>Bsa</i>I digest and purification</b><br>
 +
<ul>
 +
<li>Experiment: <br>The PCR purification was performed using the peqGOLD Cycle-pure Kit (PeqLab), modified using columns from the peqGOLD Plasmid Miniprep Kit (PeqLab)! Elution in 100 µL EB.</li>
 +
</ul>
 +
<ul>
 +
<li>Observations & Results: <br>We now seemed to have found the mistake that caused problems with the loss of DNA material in the second mutagenesis round! Miscommunication led to using the wrong kit for purification steps. We decided to use the peqGOLD Cycle-pure Kit (PeqLab) instead of the subsequent ethanol precipitation as well, as this seemed to rise fewer problems.</li>
 +
</ul>
 +
<br>
 +
<b>V08_17_2 2<sup>nd</sup> round: Ligation</b><br>
<ul>
<ul>
<li>Experiment: <br>The PCR purification was performed using the peqGOLD Cycle-pure Kit (PeqLab), modified using columns from the peqGOLD Plasmid Miniprep Kit (PeqLab)! Elution in 100 µL EB.</li>
<li>Experiment: <br>The PCR purification was performed using the peqGOLD Cycle-pure Kit (PeqLab), modified using columns from the peqGOLD Plasmid Miniprep Kit (PeqLab)! Elution in 100 µL EB.</li>

Revision as of 15:41, 26 September 2012

Deutsch  / English 

#3 Chemoreceptor Library - 16th Week

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V08_13


2nd round: Mutagenesis PCR, 1000 µL
  • Experiment:
    The PCR was set up in a volume of 1000 µL following the protocol from week 10.


V08_14


2nd round: PCR purification
  • Experiment:
    The PCR purification was performed using the peqGOLD Cycle-pure Kit (PeqLab), modified using columns from the peqGOLD Plasmid Miniprep Kit (PeqLab)! Elution in 100 µL EB.
  • Observations & Results:
    We now seemed to have found the mistake that caused problems with the loss of DNA material in the second mutagenesis round! Miscommunication led to using the wrong kit for purification steps. We decided to use the peqGOLD Cycle-pure Kit (PeqLab) instead of the subsequent ethanol precipitation as well, as this seemed to rise fewer problems.


V08_15


V08_15_1 2nd round: DpnI/BsaI digest and purification
  • Experiment:
    The digest was performed according to protocol from week 10. The subsequent purification was carried out using the peqGOLD Cycle-pure Kit (PeqLab)!!!
  • Observations & Results:
    The corresponding agarose gel showed band of the expected sizes! At last, there did not seem to be any loss of DNA material.

V08_15_2 2nd round: Ligation, 2000 µL
  • Experiment:
    The ligation was performed according to the establoshed protocol from week 10. Incubation over night at 16 °C.


V08_16


V08_16_1 2nd round: Purification of ligation from V08_15
  • Experiment:
    The purification of the ligation was now done using the peqGOLD Cycle-pure Kit (PeqLab) instead of an ethanol precipitation.
  • Observations & Results:
    We now seemed to have found the mistake that caused problems with the loss of DNA material in the second mutagenesis round! Miscommunication led to using the wrong kit for purification steps. We decided to use the peqGOLD Cycle-pure Kit (PeqLab) instead of the subsequent ethanol precipitation as well, as this seemed to rise fewer problems.

V08_16_2 2nd round: Ligation
  • Experiment:
    The PCR purification was performed using the peqGOLD Cycle-pure Kit (PeqLab), modified using columns from the peqGOLD Plasmid Miniprep Kit (PeqLab)! Elution in 100 µL EB.
  • Observations & Results:
    We now seemed to have found the mistake that caused problems with the loss of DNA material in the second mutagenesis round! Miscommunication led to using the wrong kit for purification steps. We decided to use the peqGOLD Cycle-pure Kit (PeqLab) instead of the subsequent ethanol precipitation as well, as this seemed to rise fewer problems.


V08_17


V08_17_1 2nd round: DpnI/BsaI digest and purification
  • Experiment:
    The PCR purification was performed using the peqGOLD Cycle-pure Kit (PeqLab), modified using columns from the peqGOLD Plasmid Miniprep Kit (PeqLab)! Elution in 100 µL EB.
  • Observations & Results:
    We now seemed to have found the mistake that caused problems with the loss of DNA material in the second mutagenesis round! Miscommunication led to using the wrong kit for purification steps. We decided to use the peqGOLD Cycle-pure Kit (PeqLab) instead of the subsequent ethanol precipitation as well, as this seemed to rise fewer problems.

V08_17_2 2nd round: Ligation
  • Experiment:
    The PCR purification was performed using the peqGOLD Cycle-pure Kit (PeqLab), modified using columns from the peqGOLD Plasmid Miniprep Kit (PeqLab)! Elution in 100 µL EB.
  • Observations & Results:
    We now seemed to have found the mistake that caused problems with the loss of DNA material in the second mutagenesis round! Miscommunication led to using the wrong kit for purification steps. We decided to use the peqGOLD Cycle-pure Kit (PeqLab) instead of the subsequent ethanol precipitation as well, as this seemed to rise fewer problems.


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