Team:UNICAMP-Brazil/Notebooks/September 27

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(Digestion of pTet and Double Terminator)
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*<p style=”text-align:justify;”>We selected 4 colonies from each plate containing one of yesterday's transformed biobricks, and inoculated them into liquid LB-AMP medium.</p>
*<p style=”text-align:justify;”>We selected 4 colonies from each plate containing one of yesterday's transformed biobricks, and inoculated them into liquid LB-AMP medium.</p>
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*<p style=”text-align:justify;”>Those inocula were grew for an O/N period at 37ºC (250 rpm).</p>
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*<p style=”text-align:justify;”>Those inocula were grown for an O/N period at 37ºC (250 rpm).</p>
''Marcelo''
''Marcelo''

Revision as of 21:06, 19 October 2009

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ColiGuard

Inoculation of Yesterday's Ressuspended and Transformed Biobricks

  • We selected 4 colonies from each plate containing one of yesterday's transformed biobricks, and inoculated them into liquid LB-AMP medium.

  • Those inocula were grown for an O/N period at 37ºC (250 rpm).

Marcelo

Digestion of pTet and Double Terminator

  • We pretend to ligate the parts which we are recovering right now (started yesterday) onto biobricks BBa_R0040 (pTet) and BBa_B0015 (double terminator), using it's own vector.

  • Those biobricks were already recovered on August 10th.

  • BBa_R0040 was digested with SpeI and PstI, once we intend to add a fragment in front of it (downstream). As for BBa_B0015, the digestion was realized with EcoRI and Xba restriction enzymes, once we need to insert a fragment behind it's part (upstream).

  • Digestion lasted 3 hours.

Marcelo

YeastGuard

Dephosphorylation - CIAP test

  • The E.coli didn´t grow in any of the plates. =[ We think that we didn´t plate an enough volume of bacteria. So we decided to replate a larger volume. We hope to find colonies tomorrow!

Raíssa and Taís