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| - | <li><a href="https://2010.igem.org/Team:Macquarie_Australia">Home</a></li>
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| - | <li><a href="https://2010.igem.org/Team:Macquarie_Australia/Acknowledgements">Acknowledgements</a></li>
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| - | <h1><font color="#47484c">PROJECT LAB BOOK <p>
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| - | Welcome to the Macquarie University project lab book page! <p>
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| - | Here you will find a day-by-day account of our triumphs and failures.</font></h1> </hr>
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| - | <center><h2> A day-by-day progress for <i> Agrobacterium Tumefaciens </i> Bacteriophytochrome </h2> </center>
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| - | 20th August 2010 <p>
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| - | Genomic DNA extraction <p> </big>
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| - | <menu>
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| - | <li type="disc">
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| - | <i> A. tumefaciens </i> genomic DNA extracted using the Invitrogen PureLinkTM Genomic DNA Purification kit as per the manufacturer’s protocols. </li> <li type="disc"> <p>
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| - | The extractions were run on a GelRed stained 1% agarose gel and photo taken for visualization (see below). </li> <p> <li type="disc">
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| - | A NanoDrop spectrophotometer reading was also recorded to check the quality of the extracted genomic DNA. <p> </li> <li type="disc">
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| - | The extraction was successful for all A. tumefaciens cell lysate samples (labeled DNA1.1, DNA1.2, DNA2 (See figure below) <p> </li>
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| - | <p><h3><i><b>A. tumefaciens </i> genomic DNA extraction agarose results: <p> </b> </h3>
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| - | All four DNA samples show a smear of gDNA. Because the samples were treated with RNase there is no band indicative of RNA visible = SUCCESS! <p>
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| - | <h4> <center>***** Results of A. tumefaciens genomic DNA extraction ***</h4></center></big> <p>
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| - | <b>Figure 1. </b> GelRed stained 1% agarose gel of genomic DNA extraction from A. tumefaciens.<p>
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| - | In lanes 1 and 11 there is a 1kb ladder. In lane 2 is the DNA1.1 flow through, lane 3 is the DNA2.2 flow through, lane 4 is the DNA2.1 flow through and lane 4 in the DNA2.2 flowthrough. All four samples show a smear that is indicative of genomic DNA. <u> The extraction has been successful. </u>
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| - | <h4>Nanodrop absorbance readings: </h4>
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| - | <th>Genomic DNA sample</th>
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| - | <th>260/280 OD ratio</th>
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| - | <th>Concentration (ng/mL)</th>
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| - | </tr>
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| - | <td>DNA1.1</td>
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| - | <td>1.88</td>
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| - | <td>351.5</td>
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| - | </tr>
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| - | <td>DNA1.2</td>
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| - | <td>1.98</td>
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| - | <td>203.1</td>
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| - | </tr>
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| - | <td>DNA2.1</td>
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| - | <td>2.14</td>
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| - | <td>738.9</td>
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| - | </tr>
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| - | <td>DNA2.2</td>
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| - | <td>2.16</td>
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| - | <td>709.7</td>
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| - | </tr>
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| - | </table>
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| - | </center>
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| - | <p> </div>
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| - | Overall, the Nanodrop readings show that we have obtained good DNA concentration with minimal protein contamination <p>= <u>SUCCESS! </u><p>
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| - | <big><hr>
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| - | 27th August 2010 <p>
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| - | Primer design <p> </big> </hr>
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| - | <li type="disc">
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| - | Various primers were designed manually and using Primer 3 Software package for PCR amplification.</li> <p>
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| - | <li type="disc">
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| - | The primers were ordered and supplied through XXXXXXXXXXXXXXXXX.</li><p>
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| - | <li type="disc">
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| - | There was an array of various primers ordered for amplification of different products. The details of the primers are described below. </li><p>
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PROJECT LAB BOOK
Welcome to the Macquarie University project lab book page!
Here you will find a day-by-day account of our triumphs and failures.
A day-by-day progress for Agrobacterium Tumefaciens Bacteriophytochrome
20th August 2010
Genomic DNA extraction
"
