Team:UNICAMP-Brazil/Notebooks/October 6

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*<p style=”text-align:justify;”> We did the miniprep of BBa B0015 and BBa K112806 according to [https://2009.igem.org/Team:UNICAMP-Brazil/Protocols/Mini-Prep Protocol 2]</p>
*<p style=”text-align:justify;”> We did the miniprep of BBa B0015 and BBa K112806 according to [https://2009.igem.org/Team:UNICAMP-Brazil/Protocols/Mini-Prep Protocol 2]</p>
*<p style=”text-align:justify;”> With the protduct of the miniprep we made the digestion of BBa B0015 with EcoRI and XbaI and with EcoRI and SpeI to BBa K112806. After 3 hours of digestion we purified BBa K112806 from agarose gel and BBa B0015 directly from the digestion reaction.</p>
*<p style=”text-align:justify;”> With the protduct of the miniprep we made the digestion of BBa B0015 with EcoRI and XbaI and with EcoRI and SpeI to BBa K112806. After 3 hours of digestion we purified BBa K112806 from agarose gel and BBa B0015 directly from the digestion reaction.</p>
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*<p style=”text-align:justify;”> Using the purified digestions we did the ligation of BBa B0015 with BBa K112806, [https://2009.igem.org/Team:UNICAMP-Brazil/Protocols/T4_DNA_Ligase Protocol 11], we staterd to quantify our samples with the new lab's espectrophotometer.</p>
+
*<p style=”text-align:justify;”> Using the purified digestions we did the ligation of BBa B0015 with BBa K112806 and to test the digestion of BBa B0015 we did a ligation only without BBa K112806, if colonies grow with this ligation it means BBa B0015 is recircularizing [https://2009.igem.org/Team:UNICAMP-Brazil/Protocols/T4_DNA_Ligase Protocol 11], we staterd to quantify our samples with the new lab's espectrophotometer.</p>
*<p style=”text-align:justify;”> We transformed E. coli with the ligation, we hope this time it works!!</p>
*<p style=”text-align:justify;”> We transformed E. coli with the ligation, we hope this time it works!!</p>

Revision as of 21:21, 20 October 2009

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ColiGuard

Another Miniprep, another digestion, another ligation and once again, another transformation

Transformation of the BBa K112806 + BBa B0015 ligation

  • We did the miniprep of BBa B0015 and BBa K112806 according to Protocol 2

  • With the protduct of the miniprep we made the digestion of BBa B0015 with EcoRI and XbaI and with EcoRI and SpeI to BBa K112806. After 3 hours of digestion we purified BBa K112806 from agarose gel and BBa B0015 directly from the digestion reaction.

  • Using the purified digestions we did the ligation of BBa B0015 with BBa K112806 and to test the digestion of BBa B0015 we did a ligation only without BBa K112806, if colonies grow with this ligation it means BBa B0015 is recircularizing Protocol 11, we staterd to quantify our samples with the new lab's espectrophotometer.

  • We transformed E. coli with the ligation, we hope this time it works!!

Ane and Marcos

YeastGuard

New biobricks - New strategy

- We inoculated Jen1orf-pGEM and Jen1promoter-pGEM transformed colonies in liquid media to perform the screening tomorrow.

- We need more parts!! We permormed PCR to obtain the following parts: End, Jen1 orf and promoter Jen1. Unfortunately only Jen1 orf amplified.

GEL!!

Raíssa





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