Team:UNICAMP-Brazil/Notebooks/September 26

From 2009.igem.org

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====Kamikaze System====
 
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*<p style=”text-align:justify;”>  We made the digestion of BBa K112806 with EcoRI and SpeI, BBa B0015 with EcoRI and XbaI and BBa I746911 with SpeI and Pst. </p>
 
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*<p style=”text-align:justify;”>  We purified the digestion and made the ligation of BBa K112806 in the BBa B0015 plasmid according to the [https://2009.igem.org/Team:UNICAMP-Brazil/Protocols/T4_DNA_Ligase Protocol 11] </p>
 
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''Marcos''
 
====Recovering More Biobricks====
====Recovering More Biobricks====

Revision as of 20:18, 20 October 2009

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ColiGuard

New Strategy / finO and finP

  • We are truly disappointed that we still couldn't make our finO and finP biobricks. We found a lot of issues that must be solved in order to achieve this objective, and we are totally running out of time =(

  • We decided that we are going to leave finO and finP aside for a while, and we will focus on constructing our modified version of Paris 2007 Team's biobricks.

  • We made a whole schedule for that, and we hope we would complete it in a week tops. The next step would be it's insertion into genomic DNA by homologous recombination.

Marcelo



Recovering More Biobricks

  • We started our "new strategy" (related to modifying Paris 2007 team's biobricks) by recovering some biobricks that had now became useful for our project. We ressuspended biobricks BBa_E0840 (GFP device), BBa_I718017 (lox71), BBa_J61000 (chloramphenicol resistance cassette) and BBa_I718016 (lox66), and then transfomed them into electrocompetent E. coli cells, strain DH10B, according to Protocol 3.

  • The transformed cells were plated in LB-AMP media, and were grown for an O/N period.

Marcelo and Victor

YeastGuard

Dephosphorylation - CIAP test

  • Today we performed 5' dephosphorylation of a biobrick vector (previously digested with XbaI and SpeI) with CIAP (Protocol 10) in order to test the reaction efficiency and compare with SAP protocol (Protocol 9).

  • We did two ligation reactions, one using the phosphorylated vector (control) and another using the dephosphorylated (Protocol 11).

  • We transformed the electrocompetent E. coli (Protocol 3) with the ligations and plated in LB+Amp+Kan media.

Raíssa and Taís


Today we performed a colony PCR.

Wesley and Gleidson





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