Team:UNICAMP-Brazil/Notebooks/September 24

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====''E. coli'' transformation with F plasmid====
====''E. coli'' transformation with F plasmid====
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*<p style=”text-align:justify;”>''E. coli'' DH10β electrocompetent cells were transformed using the recircularized F plasmid, according to [https://2009.igem.org/Team:UNICAMP-Brazil/Protocols/Electroporation Protocol 3].</p>
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*<p style=”text-align:justify;”>''E. coli'' DH10B electrocompetent cells were transformed using the recircularized F plasmid, according to [https://2009.igem.org/Team:UNICAMP-Brazil/Protocols/Electroporation Protocol 3].</p>
*<p style=”text-align:justify;”>After the incubation time, 100 μl of medium containing the transformed cells were plated in LB-AMP-Xgal culture plates, and incubated O/N at 37°C.</p>
*<p style=”text-align:justify;”>After the incubation time, 100 μl of medium containing the transformed cells were plated in LB-AMP-Xgal culture plates, and incubated O/N at 37°C.</p>
''Gabriel''
''Gabriel''
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====finO and finP - Still Trying to Confirm our Biobricks====
====finO and finP - Still Trying to Confirm our Biobricks====
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''Marcelo''
''Marcelo''
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==== PY Promoter - Transformation ====
 
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*<p style=”text-align:justify;”>Today we transformed the ligations PY1 + pGEM and PY2 + pGEM into electrocompetent ''E. coli'' bacteria, strain DH10B. We followed [https://2009.igem.org/Team:UNICAMP-Brazil/Protocols/Electroporation Protocol 3] (Electroporation) without modifications.</p>
 
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*<p style=”text-align:justify;”>After the transformation we plated the transformed cells in LB-AMP-Xgal plates, and let them grow at 37ºC for an O/N period.</p>
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==== CeaB and CeiB: Miniprep and transformation ====
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*<p style=”text-align:justify;”>Today, we performed miniprep ([https://2009.igem.org/Team:UNICAMP-Brazil/Protocols/Mini-prep Protocol 2]) of the transformed cells and PCR again, so that avoids unspecific amplification. We didn’t get the expected band size again.</p>
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''Fabi and Léo''
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''Luige''
{{:Team:UNICAMP-Brazil/inc_rodape}}
{{:Team:UNICAMP-Brazil/inc_rodape}}

Latest revision as of 03:10, 22 October 2009

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ColiGuard

E. coli transformation with F plasmid

  • E. coli DH10B electrocompetent cells were transformed using the recircularized F plasmid, according to Protocol 3.

  • After the incubation time, 100 μl of medium containing the transformed cells were plated in LB-AMP-Xgal culture plates, and incubated O/N at 37°C.

Gabriel

finO and finP - Still Trying to Confirm our Biobricks

  • After the miniprep procedure, we performed PCRs with finO and finP's designed primer, in order to confirm that our inserted fragment is indeed contained in the vector.

  • As we had already got an amplified fragment of expected size from those colonies, we expect that those PCRs results in a successful amplification of our fragments. Moreover, as we aimed in reducing the amount of genomic DNA, we should got less inespecific amplifications.


Marcelo


CeaB and CeiB: Miniprep and transformation

  • Today, we performed miniprep (Protocol 2) of the transformed cells and PCR again, so that avoids unspecific amplification. We didn’t get the expected band size again.


Luige




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