Team:UNICAMP-Brazil/Notebooks/September 24

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==''' ColiGuard '''==
==''' ColiGuard '''==
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====''E. coli'' transformation with F plasmid====
====''E. coli'' transformation with F plasmid====
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*''E. coli'' DH10β electrocompetent cells were transformed using the recircularized F plasmid, according to protocol 4.
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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>
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*After the incubation time, 100 μl of medium containing the transformed cells were plated in LB + ampicilin + Xgal culture plates, and incubated O/N at 37°C.
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*<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====
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*<p style=”text-align:justify;”>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.</p>
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*<p style=”text-align:justify;”>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.</p>
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''Marcelo''
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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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''Luige''
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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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