Team:UNICAMP-Brazil/Notebooks/September 26
From 2009.igem.org
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''Marcelo'' | ''Marcelo'' | ||
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====Recovering More Biobricks==== | ====Recovering More Biobricks==== | ||
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''Marcelo and Victor'' | ''Marcelo and Victor'' | ||
+ | ====Cre-Recombinase and pSB1A3 - New digestion==== | ||
+ | * Today we repeated both Cre-Recombinase's and pSB1A3's digestion with ''Xba''I and ''Spe''I. Digestion lasted 3 hours. | ||
+ | * Then, we ran an 1% agarose gel in order to confirm that digestion actually worked. | ||
+ | * Sadly, it didn't. =/ | ||
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+ | ''Víctor'' | ||
==''' YeastGuard '''== | ==''' YeastGuard '''== | ||
- | ====Dephosphorylation | + | ====Solving the recircularization problems: Dephosphorylation with CIAP==== |
+ | *<p style=”text-align:justify;”>The dephosphorylation of the vectors theoretically avoids its recircularization. We tried a diferent protocol from the one used by ColiGuard.</p> | ||
- | *<p style=”text-align:justify;”> | + | *<p style=”text-align:justify;”>We performed 5' dephosphorylation of a biobrick vector (previously digested with ''Xba''I and ''Spe''I) with CIAP ([https://2009.igem.org/Team:UNICAMP-Brazil/Protocols/CIAP_Dephosphorylation Protocol 10]) in order to test the reaction efficiency and compare with SAP protocol ([https://2009.igem.org/Team:UNICAMP-Brazil/Protocols/SAP_Dephosphorylation Protocol 9]).</p> |
*<p style=”text-align:justify;”>We did two ligation reactions, one using the phosphorylated vector (control) and another using the dephosphorylated ([https://2009.igem.org/Team:UNICAMP-Brazil/Protocols/T4_DNA_Ligase Protocol 11]).</p> | *<p style=”text-align:justify;”>We did two ligation reactions, one using the phosphorylated vector (control) and another using the dephosphorylated ([https://2009.igem.org/Team:UNICAMP-Brazil/Protocols/T4_DNA_Ligase Protocol 11]).</p> | ||
- | *<p style=”text-align:justify;”>We transformed the electrocompetent ''E. coli'' ([https://2009.igem.org/Team:UNICAMP-Brazil/Protocols/Electroporation Protocol 3]) with the ligations and plated in LB | + | *<p style=”text-align:justify;”>We transformed the electrocompetent ''E. coli'' ([https://2009.igem.org/Team:UNICAMP-Brazil/Protocols/Electroporation Protocol 3]) with the ligations and plated in LB-AMP-KAN media. </p> |
''Raíssa and Taís'' | ''Raíssa and Taís'' | ||
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{{:Team:UNICAMP-Brazil/inc_rodape}} | {{:Team:UNICAMP-Brazil/inc_rodape}} |
Latest revision as of 03:18, 22 October 2009
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