Team:SDU-Denmark/labwork/Notebook/week4

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     <span class="classred"><a href="https://2012.igem.org/Team:SDU-Denmark/labwork/Notebook/week8">8th week      </a></span>    </regulartext></td>
     <span class="classred"><a href="https://2012.igem.org/Team:SDU-Denmark/labwork/Notebook/week8">8th week      </a></span>    </regulartext></td>
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    <span class="classred"><a href="https://2012.igem.org/Team:SDU-Denmark/labwork/Notebook/week9">9th week</a></span>    </regulartext></td>
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    <span class="classred"><a href="https://2012.igem.org/Team:SDU-Denmark/labwork/Notebook/week10">10th week</a></span>    </regulartext></td>
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    <span class="classred"><a href="https://2012.igem.org/Team:SDU-Denmark/labwork/Notebook/week11">11th week</a></span>    </regulartext></td>
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    <span class="classred"><a href="https://2012.igem.org/Team:SDU-Denmark/labwork/Notebook/week12">12th week</a></span>    </regulartext></td>
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<p> <b>23-07-2012 to 29-07-2012</b> </p>
<p> <b>23-07-2012 to 29-07-2012</b> </p>
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<h2>SST send for sequencing </h2> <br/>
 
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Another checkdigest was made on SST colonies, we had our doubts about the entegrity of the formerly used restriction enzyme, so a newer EcoRI was used, a get was made and this time we got results. After a checkdigest that showed bands at expected lengths and nanodrop, two samples of 1-SST was sent for sequencing.<br/><br/>
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SST from the gel extraction the 21-07-2012 was diluted to a concentration of 100ng/µL and separated to 4 samples of 15µL each and send of for sequencing. <br/>
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4 days later we recieved results from sequencing. As we expected from the original ncbi sequences, the genes contained illegal restriction sites.<br/>
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We made liquid cultures from the SST plates from 19-07-2012 and incubated them O.N.
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1-SST contained 1 SpeI restriction site and 1-FFT had 1 EcoRI and 1 XbaI. <br/><br/>
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<h2>Cryo backup, miniprep and preparation to sequencing </h2> <br/>
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We made some Cryo backup’s from the 18 liquid cultures, that was incubated O.N. and took some sample out from each to do a miniprep on. <br/>  
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We ran a gel on the samples and found that two samples had bands corresponding to our cut gene and vector.  
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These will be shipped of to be sequneced.
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<h2> SST send for sequencing </h2> <br/>
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The two sample from earlier were prepared for sequencing by diluting them to 70-78 ng/μl and then send. Afterwards we looked into the expected outcomes of sequencing and had a look at the following mutagenesis post-sequencing.
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Primers for mutagenesis were constructed and ordered with the help of our supervisor Steffen Schmidt
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</p>

Latest revision as of 02:07, 27 September 2012

iGEM TEAM ::: SDU-DENMARK

Laboratory Notebook

1st week 2nd week 3th week 4th week
5th week 6th week 7th week 8th week
9th week 10th week 11th week 12th week

23-07-2012 to 29-07-2012

Another checkdigest was made on SST colonies, we had our doubts about the entegrity of the formerly used restriction enzyme, so a newer EcoRI was used, a get was made and this time we got results. After a checkdigest that showed bands at expected lengths and nanodrop, two samples of 1-SST was sent for sequencing.

4 days later we recieved results from sequencing. As we expected from the original ncbi sequences, the genes contained illegal restriction sites.
1-SST contained 1 SpeI restriction site and 1-FFT had 1 EcoRI and 1 XbaI.

Primers for mutagenesis were constructed and ordered with the help of our supervisor Steffen Schmidt

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