Team:UNIPV-Pavia/Calendar/July/settimana2

      <td>1</td>
      <td></html><a href="http://partsregistry.org/wiki/index.php/Part:BBa_I13501">BBa_I13501</a> (X-P)</td>
      <td>7</td>
      <td>1</td>
      <td>1</td>
  </tr>

   <tr>
      <td>E6</td>
      <td><a href="http://partsregistry.org/wiki/index.php/Part:BBa_B0031">BBa_B0031</a> (S-P)</td>
      <td>1</td>
      <td><a href="http://partsregistry.org/wiki/index.php/Part:BBa_I13501">BBa_I13501</a> (X-P)</td>
      <td>7</td>
      <td>1</td>
      <td>1</td>
  </tr>

   <tr>
      <td>E7</td>
      <td><a href="http://partsregistry.org/wiki/index.php/Part:BBa_B0032">BBa_B0032</a> (S-P)</td>
      <td>1</td>
      <td><a href="http://partsregistry.org/wiki/index.php/Part:BBa_I13501">BBa_I13501</a> (X-P)</td>
      <td>7</td>
      <td>1</td>
      <td>1</td>
  </tr>

</table> </center>

July, 6th

<a href="http://partsregistry.org/wiki/index.php/Part:BBa_C0261">BBa_C0261</a> was resuspended from iGEM 2011 kit distribution, Plate 1, well 14C in 15 μl of ddH₂O; <a href="http://partsregistry.org/wiki/index.php/Part:BBa_C0261">BBa_C0261</a>, E1, E2, E3, E4, E5, E6, E7, and E8 were transformed in 100 μl of TOP10 competent cells according to protocols. Plates were incubated ON at 37°C.
500 ml of LB without antibiotic were prepared.

July, 7th

All plates showed a lot of colonies, except for E-8 (5 colonies); two colonies for E1, E2, E3, E4, E5, E6, E7 plates and three colonies for E8 plate were picked and inoculated in 10 μl LB for inoculum and screening PCR.
A 20x mix was prepared for PCR reaction:

24 μl were aliquoted in each tube, together with 1 μl of each liquid culture. PCR cycles parameters were set as follows:

• 94°C 30 seconds (denaturing)
• 60°C 1 minute (annealing)
• 72°C 1 minute and 20 seconds (elongation)

35 cycles were performed.
Liquid cultures were then inoculated in 800 μl LB with the proper antibiotic.
A small size and a medium size agarose gel were prepared.

After the PCR, gel run was carried out on the samples:

Medium size gel

Small size gel

Except for E2-1, E4-1 and E7-1, other band lengths were correct; 750 μl of E1-2, E2-2, E3-1, E4-2, E5-2, E6-1, E7-2 and E8-3 liquid cultures were used to prepare glycerol stocks while the remaining 50 μl were refilled to 5 ml for plasmid purification and incubated at 37°C 220 rpm. BBa_R0040, BBa_C0261 and BBa_I13521 were also inoculated.
In the late afternoon the team met to talk about the wet-lab activity, the abstract and the bio-safety section.

July, 8th

Cultures were grown till saturation; plasmid purification was carried out:

• Recent changes
• What links here
• Related changes
• Special pages
• My preferences

• Printable version
• Permanent link
• Privacy policy
• Disclaimers