Team:Marburg:Project:Notebook:April
From 2014.igem.org
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<img src="https://static.igem.org/mediawiki/2014/4/42/2014-04-08_flagellin.jpg" width="15%"/> | <img src="https://static.igem.org/mediawiki/2014/4/42/2014-04-08_flagellin.jpg" width="15%"/> | ||
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<h3>Results</h3> | <h3>Results</h3> | ||
<p>Expected bands: 2kb band (whole Flank-Hag fragment)</p> | <p>Expected bands: 2kb band (whole Flank-Hag fragment)</p> | ||
- | <img src="https://static.igem.org/mediawiki/2014/8/81/2014-04-10_Hag_Flank2.jpg" width=" | + | <img src="https://static.igem.org/mediawiki/2014/8/81/2014-04-10_Hag_Flank2.jpg" width="15%"/> |
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<li>Incubate at 30°C till monday</li> | <li>Incubate at 30°C till monday</li> | ||
</ul> | </ul> | ||
- | <img src="https://static.igem.org/mediawiki/2014/a/ab/2014-04-10_Hag_Flank3.jpg" width=" | + | <img src="https://static.igem.org/mediawiki/2014/a/ab/2014-04-10_Hag_Flank3.jpg" width="15%" /> |
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Lane 1 & 2 contain the expected Fragments (~ 600 bp). In Lane 3 there is a very thin fragment with the expected size (~ 1100 bp) but smaller fragments can also be seen. Lane 5 does not contain any fragment at all and in lane 6 there is the fragment of the expected size (~ 600 bp). Both samples with fragment 1 were pooled, sample 3.1 was discarded and the whole PCR-product 2.1 has been separated in an agarosegel and the fragment with 1100 bp has been cut out and purified with the Gel Extraction Kit (c = 4,4 ng/µl). A second PCR for fragment 2 has been done in parallel. All parameters remained the same but the annealing-temperature has been changed to 62°C.</p> | Lane 1 & 2 contain the expected Fragments (~ 600 bp). In Lane 3 there is a very thin fragment with the expected size (~ 1100 bp) but smaller fragments can also be seen. Lane 5 does not contain any fragment at all and in lane 6 there is the fragment of the expected size (~ 600 bp). Both samples with fragment 1 were pooled, sample 3.1 was discarded and the whole PCR-product 2.1 has been separated in an agarosegel and the fragment with 1100 bp has been cut out and purified with the Gel Extraction Kit (c = 4,4 ng/µl). A second PCR for fragment 2 has been done in parallel. All parameters remained the same but the annealing-temperature has been changed to 62°C.</p> | ||
- | <img src="https://static.igem.org/mediawiki/2014/a/a0/2014-04-15_mutagenesis2.jpg" width=" | + | <img src="https://static.igem.org/mediawiki/2014/a/a0/2014-04-15_mutagenesis2.jpg" width="20%" /> |
<p><strong>Figure : modified mutagenesis PCR for fragment 2; again there are two fragments with a too small size and the expected fragments are not high concentrated.</strong></p> | <p><strong>Figure : modified mutagenesis PCR for fragment 2; again there are two fragments with a too small size and the expected fragments are not high concentrated.</strong></p> | ||
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<p>To be sure to obtain just a fragment of the wanted size a PCR with the same Primers (iGEM-003 and iGEM-010) has been carried out with the thin fragment which was cut out of the gel yesterday.</p> | <p>To be sure to obtain just a fragment of the wanted size a PCR with the same Primers (iGEM-003 and iGEM-010) has been carried out with the thin fragment which was cut out of the gel yesterday.</p> | ||
- | <img src="https://static.igem.org/mediawiki/2014/f/fe/2014-04-16_gel.jpg" width=" | + | <img src="https://static.igem.org/mediawiki/2014/f/fe/2014-04-16_gel.jpg" width="15%" /> |
<p><strong>Figure : PCR of the fragment 2; <br /> | <p><strong>Figure : PCR of the fragment 2; <br /> | ||
1: 2-log-DNA-ladder, <br /> | 1: 2-log-DNA-ladder, <br /> | ||
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<legend><a name="exp15.9">15.9 PCR-Amplification of Flagellin constructs</a></legend> | <legend><a name="exp15.9">15.9 PCR-Amplification of Flagellin constructs</a></legend> | ||
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<div class="results"> | <div class="results"> | ||
<h3>Results</h3> | <h3>Results</h3> | ||
- | <img src="https://static.igem.org/mediawiki/2014/4/42/2014-04-23_flagellin.jpg" width=" | + | <img src="https://static.igem.org/mediawiki/2014/4/42/2014-04-23_flagellin.jpg" width="20%"/> |
<p>Figure: PCR of the Hag-Flank-construct</p> | <p>Figure: PCR of the Hag-Flank-construct</p> | ||
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Revision as of 16:18, 12 October 2014
Notebook: April