Difference between revisions of "Team:Pasteur Paris/Week 13"
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− | <h3 | + | <center><h3><em>Yeast assembly</em></h3></center></br> |
<ul style="text-align: left"> | <ul style="text-align: left"> | ||
<li>Treatment of the yeast using a homemade Zymolyase solution :</li> | <li>Treatment of the yeast using a homemade Zymolyase solution :</li> | ||
<ol> | <ol> | ||
− | <li> | + | <li> PBS pH 7.5 5 ml</li> |
<li>glucose 5% to make a 2% solution</li> | <li>glucose 5% to make a 2% solution</li> | ||
− | <li>Zymolyase 0 | + | <li>Zymolyase 0.1 g (0.012-0.022 mg/mL)</li> |
− | <li>Distilled water free RNAs/DNAs | + | <li>Distilled water free RNAs/DNAs 1 mL</li> |
<li>MidiPrep of pRS415</li> | <li>MidiPrep of pRS415</li> | ||
</ol> | </ol> | ||
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<tr> | <tr> | ||
<td width="86" valign="top"><p>pRS415</p></td> | <td width="86" valign="top"><p>pRS415</p></td> | ||
− | <td width="86" valign="top"><p align="right">6 | + | <td width="86" valign="top"><p align="right">6.2 </p></td> |
− | <td width="86" valign="top"><p align="right">0 | + | <td width="86" valign="top"><p align="right">0.122 </p></td> |
− | <td width="86" valign="top"><p align="right">0 | + | <td width="86" valign="top"><p align="right">0.067 </p></td> |
− | <td width="86" valign="top"><p align="right">0 | + | <td width="86" valign="top"><p align="right">0.076 </p></td> |
− | <td width="86" valign="top"><p align="right">1 | + | <td width="86" valign="top"><p align="right">1.60 </p></td> |
− | <td width="86" valign="top"><p align="right">1 | + | <td width="86" valign="top"><p align="right">1.83 </p></td> |
</tr> | </tr> | ||
</table> | </table> | ||
+ | |||
<p style="text-align: left">MidiPrep has been digested to verified if the DNA correspond to the plasmid of interest or to an other plasmid that is naturally present in yeast. </p> | <p style="text-align: left">MidiPrep has been digested to verified if the DNA correspond to the plasmid of interest or to an other plasmid that is naturally present in yeast. </p> | ||
<ul style="text-align: left"> | <ul style="text-align: left"> | ||
<li>BamHI digestion of pRS415 (with the phosphatase step). </li> | <li>BamHI digestion of pRS415 (with the phosphatase step). </li> | ||
</ul> | </ul> | ||
− | <p style="text-align: left">We can see the presence of a streak about | + | <p style="text-align: left">We can see the presence of a streak about 7500 bp which seems to correspond to our plasmid pRS415.</p> |
<ul style="text-align: left"> | <ul style="text-align: left"> | ||
<li>DNA concentration measurement. </li> | <li>DNA concentration measurement. </li> | ||
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<tr> | <tr> | ||
<td width="121" valign="top"><p>BBa_J61047</p></td> | <td width="121" valign="top"><p>BBa_J61047</p></td> | ||
− | <td width="121" valign="top"><p align="right">0 | + | <td width="121" valign="top"><p align="right">0.4 </p></td> |
− | <td width="121" valign="top"><p align="right">0 | + | <td width="121" valign="top"><p align="right">0.008 </p></td> |
− | <td width="121" valign="top"><p align="right">0 | + | <td width="121" valign="top"><p align="right">0.002 </p></td> |
− | <td width="121" valign="top"><p align="right">3 | + | <td width="121" valign="top"><p align="right">3.76 </p></td> |
</tr> | </tr> | ||
</table> | </table> | ||
+ | |||
<ul style="text-align: left"> | <ul style="text-align: left"> | ||
<li>PCR amplification of pRS415 with a mutagenesis kit using TE 8.1. </li> | <li>PCR amplification of pRS415 with a mutagenesis kit using TE 8.1. </li> | ||
Line 175: | Line 177: | ||
</ul> | </ul> | ||
<p> </p> | <p> </p> | ||
− | <h3 | + | <center><h3><em>TPA solubility</em></h3></center></br> |
<ul style="text-align: left"> | <ul style="text-align: left"> | ||
<li>improvement of TPA solubility in different conditions : </li> | <li>improvement of TPA solubility in different conditions : </li> | ||
<ol> | <ol> | ||
− | + | <li>TPA (0.1 g) + Na<sub>2</sub>H<sub>2</sub>PO<sub>4</sub>(0.5 g) in 150 ml of water: complete TPA solubilization. </li> | |
− | <li>TPA (0. | + | <li>TPA (0.1 g) + NaOH(0.5 g) in 150 ml of water: complete TPA solubilization. </li> |
− | <li>TPA (0. | + | |
− | + | ||
</ol> | </ol> | ||
<li>Fluorescence measurements : </li> | <li>Fluorescence measurements : </li> |
Latest revision as of 23:47, 3 October 2015
Week 1Week 2Week 3Week 4Week 5Week 6Week 7Week 8Week 9Week 10Week 11Week 12Week 13Week 14Week 15 |
Week 13 Yeast assembly
MidiPrep has been digested to verified if the DNA correspond to the plasmid of interest or to an other plasmid that is naturally present in yeast.
We can see the presence of a streak about 7500 bp which seems to correspond to our plasmid pRS415.
TPA solubility
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