Team:UNICAMP-Brazil/Notebooks/September 27

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====Digestion of pTet and Double Terminator====
====Digestion of pTet and Double Terminator====
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*<p style=”text-align:justify;”>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.</p>
+
*<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>
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*<p style=”text-align:justify;”>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).</p>
+
*<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====
====CeaB and CeiB====
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*<p style=”text-align:justify;”>We don’t have time, the deadline is next. So, we decided just isolate both CeaB and CeiB in a plasmid backbone. We chose isolating into pSB1A3 plasmid. Thus, we digested the plasmid with the right restriction enzymes (SpeI and XbaI). Immediately, we made the purification gel according to the [https://2009.igem.org/Team:UNICAMP-Brazil/Protocols/Purification_of_DNA_fragments_from_agarose_gels  protocol 7].</p>
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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.
 +
 +
''Víctor''
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''Luige''
 
==''' 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>
 +
 +
* 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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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 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.

  • 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 XbaI restriction enzymes, once we need to insert a fragment behind it's part (upstream).

  • Digestion lasted 3 hours.

Marcelo

CeaB and CeiB

  • 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 (SpeI and XbaI). Immediately, we made the purification gel according to Protocol 7.

Luige

Cre-Recombinase - New PCR

  • 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.

Víctor


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!

  • Meanwhile we kept screening the old transformations. No sucsess. =(

Raíssa and Taís




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