Team:Exeter/lab book/gibs/wk5

From 2012.igem.org

(Difference between revisions)
 
Line 160: Line 160:
   
   
  </table>
  </table>
 +
 +
<table width="980" align="center" cellspacing="20">
 +
<tr align="center">
 +
  <td>
 +
  <font color="#57B947" size="1" face="Verdana">
 +
    <p><u>Website Designed and Built by: Ryan Edginton, James Lynch & Alex Clowsley</u> &nbsp;&nbsp;|&nbsp;&nbsp;
 +
    <a href="https://igem.org/Team.cgi?id=764" style="color:#57B947" target="_blank"><u>Contact Us</u></a>  &nbsp;&nbsp;|&nbsp;&nbsp;
 +
    <a href="https://2012.igem.org/Team:Exeter/site_map" style="color:#57B947"><u>Site Map</u></a></p>
 +
  </font>
 +
  </td>
 +
</tr>
 +
</table>
</body>
</body>
</html>
</html>

Latest revision as of 00:10, 27 September 2012

ExiGEM2012 Lab Book Gibs wk5

Operon Construction: 6th - 10th August 2012

 **Monday 6.8.12**


3A BioBrick assembly of RBS-wbbC(d) (BBa_B0034 + BBa_K764005)


Upstream

RBS 3 (BBa_B0034) plasmid DNA - 8.1 µl

Water - 14.4 µl


Downstream wbbC(d)

DNA - 2.8 µl

Water - 19.7 µl


Destination plasmid pSB1C3 (chloramphenicol resistant) Digestion

Ligation


Upstream: 2 µl

Downstream: 2 µl

MilliQ water 2.5


23 °C (room temp.) incubation for 1 hour

20 minute heat kill 80 °C


**Wednesday 8.8.12**


Transformed RBS-wbbC(d) (BBa_B0034 + BBa_K764005)

2 µl into 25 µl competent cells

50 µl and 150 µl LB(chlor) spread plates made


**Thursday 9.8.12**


Transferred RBS-wbbC(d) (BBa_B0034 + BBa_K764005) transformed colonies to LB(chlor) medium


**Friday 10.8.12**


RBS-wbbC(d) transformed cells broth tubes centrifuged for 5 minutes, 3901 rcf, 4 °C

Supernatant discarded

Miniprep according to Fermentas protocol

Step 1045µl milli-Q water added in place of elution buffer, followed by another 5 µl after incubation and centrifugation

Website Designed and Built by: Ryan Edginton, James Lynch & Alex Clowsley   |   Contact Us   |   Site Map

Retrieved from "http://2012.igem.org/Team:Exeter/lab_book/gibs/wk5"