Team:UNICAMP-Brazil/Notebooks/October 8

From 2009.igem.org

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==''' YeastGuard '''==
==''' YeastGuard '''==
====New strategy: pGEM====
====New strategy: pGEM====
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*<p style=”text-align:justify;”>We did miniprep of pDLD-pGEM, pJEN1-pGEM and Lysozyme-pGEM. We digested these plasmids with ‘’EcoR’’I and ‘’Spe’’I to release the parts from the pGEM. All of the digestions worked so we ligated them with biofusion vector previously digested with ''EcoR''I and ''Spe''I. We transformed competent ''E. coli'' with the promoters+biofusion ligation reaction and plated in LB Amp. The lisozyme ligation reaction was performed late at night, so we left it O/N to transform tomorrow.</p>
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*<p style=”text-align:justify;”>We did miniprep of pDLD-pGEM, pJEN1-pGEM and Lysozyme-pGEM. We digested these plasmids with ''EcoR''I and ''Spe''I to release the parts from the pGEM. All of the digestions worked so we ligated them with biofusion vector previously digested with ''EcoR''I and ''Spe''I. We transformed competent ''E. coli'' with the promoters+biofusion ligation reaction and plated in LB+Amp media. The lysozyme ligation reaction was performed late at night, so we left it O/N to transform tomorrow.</p>
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GEL
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GEL
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Revision as of 01:17, 21 October 2009

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ColiGuard

Another try for Kamikaze System

  • We purify from agarose gel the digestion that we let doing yesterday and prepare another digestion with XbaI, this digestion was purified and used in a new ligation of BBa B0015 BBa K112806.

  • The ligation was used in a transformation, is the third time we do the same ligation, we hope this time we have luck!!!

Ane and Marcos


YeastGuard

New strategy: pGEM

  • We did miniprep of pDLD-pGEM, pJEN1-pGEM and Lysozyme-pGEM. We digested these plasmids with EcoRI and SpeI to release the parts from the pGEM. All of the digestions worked so we ligated them with biofusion vector previously digested with EcoRI and SpeI. We transformed competent E. coli with the promoters+biofusion ligation reaction and plated in LB+Amp media. The lysozyme ligation reaction was performed late at night, so we left it O/N to transform tomorrow.

GEL GEL


Raíssa