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Team:UNICAMP-Brazil/Notebooks/September 27
From 2009.igem.org
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====Inoculation of Yesterday's Ressuspended and Transformed Biobricks==== | ====Inoculation of Yesterday's Ressuspended and Transformed Biobricks==== | ||
| - | *<p style=”text-align:justify;”>We selected 4 colonies from each plate containing one of yesterday's transformed biobricks, and inoculated them | + | *<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;”>Those inocula were | + | *<p style=”text-align:justify;”>Those inocula were grown for an O/N period at 37ºC (250 rpm).</p> |
''Marcelo'' | ''Marcelo'' | ||
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====Digestion of pTet and Double Terminator==== | ====Digestion of pTet and Double Terminator==== | ||
| - | *<p style=”text-align:justify;”>We | + | *<p style=”text-align:justify;”>We intend 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.</p> |
*<p style=”text-align:justify;”>Those biobricks were already recovered on August 10th.</p> | *<p style=”text-align:justify;”>Those biobricks were already recovered on August 10th.</p> | ||
| - | *<p style=”text-align:justify;”>BBa_R0040 was digested with | + | *<p style=”text-align:justify;”>BBa_R0040 was digested with ''Spe''I and ''Pst''I, once we intend to add a fragment in front of it (downstream). As for BBa_B0015, the digestion was realized with ''EcoR''I and ''Xba''I restriction enzymes, once we need to insert a fragment behind it's part (upstream).</p> |
*<p style=”text-align:justify;”>Digestion lasted 3 hours.</p> | *<p style=”text-align:justify;”>Digestion lasted 3 hours.</p> | ||
''Marcelo'' | ''Marcelo'' | ||
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| + | ====CeaB and CeiB==== | ||
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| + | *<p style=”text-align:justify;”>We don’t have time, the deadline is near. So, we decided to just isolate both CeaB and CeiB in a plasmid backbone. We chose to isolate into [http://partsregistry.org/Part:pSB1A3 pSBA3] plasmid. Thus, we digested the plasmid with the right appropriate enzymes (''Spe''I and ''Xba''I). Immediately, we made the purification gel according to [https://2009.igem.org/Team:UNICAMP-Brazil/Protocols/Purification_of_DNA_fragments_from_agarose_gels Protocol 7].</p> | ||
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| + | ''Luige'' | ||
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| + | ====Cre-Recombinase - New PCR==== | ||
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| + | * After several attempts on correctly digesting Cre-Recombinase's fragment, we ended up without any sample! =/ | ||
| + | * Therefore, today we repeated the PCR reaction for amplifying Cre-Recombinase without ATG, in order to generate more sample. | ||
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| + | ''Víctor'' | ||
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==''' YeastGuard '''== | ==''' YeastGuard '''== | ||
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*<p style=”text-align:justify;”>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!</p> | *<p style=”text-align:justify;”>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!</p> | ||
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| + | * Meanwhile we kept screening the old transformations. No sucsess. =( | ||
''Raíssa and Taís'' | ''Raíssa and Taís'' | ||
{{:Team:UNICAMP-Brazil/inc_rodape}} | {{:Team:UNICAMP-Brazil/inc_rodape}} | ||
Latest revision as of 03:23, 22 October 2009
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