Team:UNICAMP-Brazil/Notebooks/September 25

From 2009.igem.org

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==''' ColiGuard '''==
==''' ColiGuard '''==
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====Kamikaze System====
 
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*<p style=”text-align:justify;”>We did the miniprep with the innoculum made yesterday using the [https://2009.igem.org/Team:UNICAMP-Brazil/Protocols/mini-prep Protocol 2]. </p>
 
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''Marcos''
 
====Getting the visa====
====Getting the visa====

Revision as of 20:18, 20 October 2009

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ColiGuard

Getting the visa

  • Today was a happy day to five of us! We went to the U.S. Embassy in Sao Paulo and we got our visas! :D

finO and finP - Still Trying to Confirm our Biobricks

  • We ran an agarose gel of yesterday's PCRs product.

  • We couldn't obtain even a single amplified fragment! =(

  • Why can the transformed cells grew in the media (that means they actually have the AMP resistence), but they haven't our inserts into it's plasmids?

  • Our advisors suggested us that, since we digested our plasmid vector with XbaI and SpeI (X sticky ends can come together with S sticky ends), it recircularized without the introduction of our inserts.

  • Therefore, we must perform the dephosphorylation of our digested vector. That may prevent it from recircularizing without the insert introduction.

Marcelo

YeastGuard

Biofusion vector - Electroelution

  • Considering that we need more vector to perform the ligation reactions, we electroeluted more vector from the ADH1 biobrick previously digested with XbaI and SpeI (Protocol 12).

Taís