Team:UNICAMP-Brazil/Notebooks/September 27

From 2009.igem.org

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''Marcelo''
''Marcelo''
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====Transformation of the BBa K112806 + BBa B0015 ligation====
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*<p style=”text-align:justify;”>We transformed the ligation made yesterday by eletroporation according to the [https://2009.igem.org/Team:UNICAMP-Brazil/Protocols/Electroporation  Protocol 3]</p>
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''Léo''
==''' YeastGuard '''==
==''' YeastGuard '''==

Revision as of 20:13, 20 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

Transformation of the BBa K112806 + BBa B0015 ligation

  • We transformed the ligation made yesterday by eletroporation according to the Protocol 3

Léo

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