|
AUGUST: WEEK 5
August, 29th
T9002-ENTERo and ENTERO-RBS were diluted 1:500 in M9 + Amp + Cm12.5 13 ml and 1 ml volume respectively. After six hour growth they were aliquoted in 96-well microplate (190 μl cultures) with 10 μl of supernatants collected on August, 27th and 10 μl of 3OC6-HSL.
Plates incubated on August, 27th were all grown, except for J101-31-1C3; for each of them a colony was picked and inoculated in LB + Cm34, 5 ml, while for J101-E7 two colonies were inoculated in LB + Cm34, 5 ml.
August, 30th
Glycerol stocks were prepared for E2-1C3, E3-1C3, E4-1C3, E5-1C3, E6-1C3, E7-1C3, E9-1C3, E10-1C3, E11-1C3, J101-E5-1C3, J101-E7-1C3-1 and J101-E7-1C3-2.
Each sample was digested with EcoRI and PstI endonucleases (a mix 13 x was prepared for high copy number plasmid parts):
while for E43-2 10 μl were digested. In the afternoon the parts were screened by gel electrophoresis:
All parts showed the correct insert length except for E3-1C3. We decided to pick and inoculate another colony from the plate, named E3-1C3-2.
August, 31st
E2-1C3, E4-1C3, E5-1C3, E6-1C3, E7-1C3, E9-1C3, E10-1C3, E11-1C3, J101-E5-1C3, J101-E7-1C3-1, E43-2 and E43-3 DNA samples were prepared and sent to BMR Genomics for sequencing.
These plasmids were digested with EcoRI and PstI endonucleases to transfer them in pSB1C3:
In gel electrophoresis all inserts showed the correct length: After gel-extraction digested DNA was quantified:
Then ligations were performed in a final volume of 10 μl:
|
Team:UNIPV-Pavia/Calendar/August/week5
From 2011.igem.org
(Difference between revisions)
(5 intermediate revisions not shown) | |||
Line 5: | Line 5: | ||
</h2> | </h2> | ||
<div class="cleared"></div> | <div class="cleared"></div> | ||
- | <div class="art-postcontent"> | + | <div class="art-postcontent" style="margin-right: 10pt;"> |
<p style="text-align:left;"> | <p style="text-align:left;"> | ||
Line 11: | Line 11: | ||
<p><a name="indice"/> | <p><a name="indice"/> | ||
</p> | </p> | ||
+ | |||
+ | <div align="justify"> | ||
<a name="August.2C_29th"></a><h2> <span class="mw-headline">August, 29th</span></h2> | <a name="August.2C_29th"></a><h2> <span class="mw-headline">August, 29th</span></h2> | ||
<p> | <p> | ||
Line 33: | Line 35: | ||
<p> | <p> | ||
- | Glycerol stocks were prepared for E2-1C3, E3-1C3, E4-1C3, E5-1C3, E6-1C3, E7-1C3, E9-1C3, E10-1C3, E11-1C3, J101-E5-1C3, J101-E7-1C3-1 and J101- | + | Glycerol stocks were prepared for E2-1C3, E3-1C3, E4-1C3, E5-1C3, E6-1C3, E7-1C3, E9-1C3, E10-1C3, E11-1C3, J101-E5-1C3, J101-E7-1C3-1 and J101-E7-1C3-2. |
<br> | <br> | ||
DNA of each culture was purified: | DNA of each culture was purified: | ||
Line 315: | Line 317: | ||
<tr> | <tr> | ||
- | <td class="row"><b> | + | <td class="row"><b>E3N-1C3</b></td> |
<td class="row"><a href="http://partsregistry.org/wiki/index.php/Part:pSB1C3">pSB1C3</a> (E-P)</td> | <td class="row"><a href="http://partsregistry.org/wiki/index.php/Part:pSB1C3">pSB1C3</a> (E-P)</td> | ||
<td class="row">1</td> | <td class="row">1</td> | ||
Line 326: | Line 328: | ||
<tr> | <tr> | ||
- | <td class="row"><b> | + | <td class="row"><b>J101-31N-1C3</b></td> |
<td class="row"><a href="http://partsregistry.org/wiki/index.php/Part:pSB1C3">pSB1C3</a> (E-P)</td> | <td class="row"><a href="http://partsregistry.org/wiki/index.php/Part:pSB1C3">pSB1C3</a> (E-P)</td> | ||
<td class="row">1</td> | <td class="row">1</td> | ||
Line 342: | Line 344: | ||
<div align="right"><small><a href="#indice" title="">^top</a></small></div> | <div align="right"><small><a href="#indice" title="">^top</a></small></div> | ||
- | + | <br> | |
<div> | <div> | ||
<span style="float:left;"> | <span style="float:left;"> | ||
Line 353: | Line 355: | ||
</span> | </span> | ||
</div> | </div> | ||
- | + | </div> | |
</html> | </html> | ||
{{endcalendar}} | {{endcalendar}} |
Latest revision as of 10:12, 18 September 2011