Team:UNICAMP-Brazil/Notebooks/September 28

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(New page: {{:Team:UNICAMP-Brazil/inc_topo}} {{:Team:UNICAMP-Brazil/inc calendar}} __NOTOC__ ==''' YeastGuard '''== ====Dephosphorylation - CIAP test==== *The ''E.coli'' didn´t grow in any of the...)
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{{:Team:UNICAMP-Brazil/inc calendar}}
{{:Team:UNICAMP-Brazil/inc calendar}}
__NOTOC__
__NOTOC__
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==''' ColiGuard '''==
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====Minipreps from BBa_E0840, BBa_I718017, BBa_J61000 and BBa_I78016====
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*<p style=”text-align:justify;”>Today we performed minipreps from yesterday's inoculated cultures, related to the biobricks already cited.</p>
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*<p style=”text-align:justify;”>We followed [https://2009.igem.org/Team:UNICAMP-Brazil/Protocols/Mini-Prep Protocol 2], without modification.</p>
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''Marcelo''
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====Purification of pTet and double terminator biobricks====
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*<p style=”text-align:justify;”>Once we digested those biobricks yesterday, we could proceed today to the purification of the target band from an agarose gel.</p>
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*<p style=”text-align:justify;”>We used Invitrogen's Purelink Quick Gel Extraction Kit, following the manufacturer's protocol ([https://2009.igem.org/Team:UNICAMP-Brazil/Protocols/Purification_of_DNA_fragments_from_agarose_gels Protocol 7]), without modifications.</p>
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''Marcelo''
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==== CeaB and CeiB transformation into plasmid [http://partsregistry.org/Part:pSB1A3 pSBA3] ====
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*<p style=”text-align:justify;”>Today, we dephosphorilated the plasmid [http://partsregistry.org/Part:pSB1A3 pSBA3], and immediately started the ligation reaction of this unphosphorylated plasmid with CeaB and CeiB. Considering that the dephosphorylation use buffer that contains salt and that salt could be interfering in the transformations, we performed a dyalisis to get the samples free of salts. At last, we transformed competent ''E. coli'' (according to the [https://2009.igem.org/Team:UNICAMP-Brazil/Protocols/Electroporation protocol 3]) and plated the respective plates. This time we expect to obtain transforming cells.</p>
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''Luige''
==''' YeastGuard '''==
==''' YeastGuard '''==
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====Dephosphorylation - CIAP test====
====Dephosphorylation - CIAP test====
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*The ''E.coli'' didn´t grow in any of the plates again. =[  We discovered that the electrocompetent ''E.coli'' that we used yesterday were dead! So we transformed another electrocompetent ''E.coli'' using the rest of the ligation reactions we had.
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*<p style=”text-align:justify;”>The ''E.coli'' didn´t grow in any of the plates again. =[  We discovered that the electrocompetent ''E.coli'' that we used yesterday were dead! So we transformed another electrocompetent ''E.coli'' using the rest of the ligation reactions we had. We extremely hope to find colonies tomorrow!</p>
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We extremely hope to find colonies tomorrow!
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''Raíssa and Taís''
''Raíssa and Taís''
{{:Team:UNICAMP-Brazil/inc_rodape}}
{{:Team:UNICAMP-Brazil/inc_rodape}}

Latest revision as of 03:26, 22 October 2009

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ColiGuard

Minipreps from BBa_E0840, BBa_I718017, BBa_J61000 and BBa_I78016

  • Today we performed minipreps from yesterday's inoculated cultures, related to the biobricks already cited.

  • We followed Protocol 2, without modification.

Marcelo


Purification of pTet and double terminator biobricks

  • Once we digested those biobricks yesterday, we could proceed today to the purification of the target band from an agarose gel.

  • We used Invitrogen's Purelink Quick Gel Extraction Kit, following the manufacturer's protocol (Protocol 7), without modifications.

Marcelo


CeaB and CeiB transformation into plasmid [http://partsregistry.org/Part:pSB1A3 pSBA3]

  • Today, we dephosphorilated the plasmid [http://partsregistry.org/Part:pSB1A3 pSBA3], and immediately started the ligation reaction of this unphosphorylated plasmid with CeaB and CeiB. Considering that the dephosphorylation use buffer that contains salt and that salt could be interfering in the transformations, we performed a dyalisis to get the samples free of salts. At last, we transformed competent E. coli (according to the protocol 3) and plated the respective plates. This time we expect to obtain transforming cells.

Luige

YeastGuard

Dephosphorylation - CIAP test

  • The E.coli didn´t grow in any of the plates again. =[ We discovered that the electrocompetent E.coli that we used yesterday were dead! So we transformed another electrocompetent E.coli using the rest of the ligation reactions we had. We extremely hope to find colonies tomorrow!

Raíssa and Taís




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