Team:SDU-Denmark/labwork/Notebook/week3

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<h2>Laboratory Notebook</h2>
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<h1>Laboratory Notebook</h1>
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Here you find the log book for the procedures carried out in the laboratory, starting from week 27. 
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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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                                             <!----------2ND WEEK---------->
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                                             <!----------3rd WEEK---------->
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<p> <b>16-07-2012 to 22-07-2012</b> </p>
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We still had no 1-SST colonies. Another PCR was made from the original cDNA, using various temperature steps. The pJET vector and 1-SST gene were run on the same gel and purified together on the same spincolumn during gel DNA extraction and mixed the elution buffer with ligation buffer and excess ligase in the final elution step.<br/><br/>
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The 1-SST colonies were incubated at 37C overnight on ampicillin agar medium plates.<br/><br/>
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A checkdigest was made on the 10 1-FFT liquid cultures with EcoRI and PstI. The resulting gel had two bands of the expected lengths for vector and insert. These liquid cultures were miniprepped and nanodropped:<br/>
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Tube#3: 138,2ng/µl<br/>
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Tube#9: 71,8ng/µl <br/><br/>
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Tube 3 contained a concentration suitable for sequencing. It was prepared and sent. We used the MWG sites primer design tool (http://www.eurofinsdna.com/) to design primers that anneal to the FFT gene. Seeing as the FFT gene is approximately 1800bp long the primers were designed to anneal between positions 450-500 and 950-1000 both in the forward direction, to be sure that the entire gene is sequenced.<br/><br/>
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The following day 1-SST cultures finaly showed some results, 6 colonies. They were transfered to liquid medium and incubated overnight.<br<br/>/>
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The 6 colonies were miniprepped and nanodropped. They all showed concentrations of >60 ng/uL, some even around 160.<br/><br/>
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The following checkdigest showed no usable results.<br/>
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Latest revision as of 02:06, 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

16-07-2012 to 22-07-2012

We still had no 1-SST colonies. Another PCR was made from the original cDNA, using various temperature steps. The pJET vector and 1-SST gene were run on the same gel and purified together on the same spincolumn during gel DNA extraction and mixed the elution buffer with ligation buffer and excess ligase in the final elution step.

The 1-SST colonies were incubated at 37C overnight on ampicillin agar medium plates.

A checkdigest was made on the 10 1-FFT liquid cultures with EcoRI and PstI. The resulting gel had two bands of the expected lengths for vector and insert. These liquid cultures were miniprepped and nanodropped:
Tube#3: 138,2ng/µl
Tube#9: 71,8ng/µl

Tube 3 contained a concentration suitable for sequencing. It was prepared and sent. We used the MWG sites primer design tool (http://www.eurofinsdna.com/) to design primers that anneal to the FFT gene. Seeing as the FFT gene is approximately 1800bp long the primers were designed to anneal between positions 450-500 and 950-1000 both in the forward direction, to be sure that the entire gene is sequenced.

The following day 1-SST cultures finaly showed some results, 6 colonies. They were transfered to liquid medium and incubated overnight./> The 6 colonies were miniprepped and nanodropped. They all showed concentrations of >60 ng/uL, some even around 160.

The following checkdigest showed no usable results.

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