Team:UNAM Genomics Mexico/prueba

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<center><h1>'''Bacillus subtilis Results'''</h1></center>  
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<p class='captionInside'>1.ArsR-CzrA 97 E/P. <br />
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2.ArsR-CzrA 98 E/P. <br />
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3.ArsR-CzrA 99 E/P. <br />
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4.CI 02 E/P. <br />
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5.P4 PCR. <br />
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We know that everybody has wetlab problems, just like us. And we actually spend most of the summer looking to standardize the Bacillus subtilis transformation protocol. And we did it! So we are presenting you our new standard:
 
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[[Team:UNAM_Genomics_Mexico/Notebook/Protocols#Escherichia_coli_MC1061_competent_cells_protocol | '''Escherichia coli MC1061 competent cells protocol''']] Please click this link to see the complete protocol in our notebook protocols
 
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[[Team:UNAM_Genomics_Mexico/Notebook/Protocols#Escherichia_coli_MC1061_heat_shock_transformation_protocol | '''Two-step Bacillus subtilis transformation procedure''']]Please click this link to see the complete protocol in our notebook protocols
 
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<h2>05/29/2012</h2><br />
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Our group is in charge of building part of the “and” construction. We started analyzing if the plasmid we have with P4 actually had what we needed. <br /><br />
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The plasmid PRMn25 contains the protein P4. It has Amp100  resistance and comes in ''Escherichia coli''  NFI. Cells were lysed to make sure the plasmid was present in these cells (5000bp). We ran a gel with 3 lysis, a sample from the plasmid PFRC54 (A3 promoter) and a sample of total DNA from the strain from which PFRC54 was obtained. <br /><br />
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We notice that you not only need a plasmid to transform into B. subtilis, but a plasmid that has form multimers in a RecA+ E. coli and should be flanked by AmyE 5' & 3', so that when the plasmid is transformed into Bacillus, it does not get degradated, and it makes an integration into the genome.
 
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Revision as of 22:50, 18 October 2012


UNAM-Genomics_Mexico

=MAY=

  • 1.ArsR-CzrA 97 E/P.
    2.ArsR-CzrA 98 E/P.
    3.ArsR-CzrA 99 E/P.
    4.CI 02 E/P.
    5.P4 PCR.










05/29/2012


Our group is in charge of building part of the “and” construction. We started analyzing if the plasmid we have with P4 actually had what we needed.

The plasmid PRMn25 contains the protein P4. It has Amp100 resistance and comes in Escherichia coli NFI. Cells were lysed to make sure the plasmid was present in these cells (5000bp). We ran a gel with 3 lysis, a sample from the plasmid PFRC54 (A3 promoter) and a sample of total DNA from the strain from which PFRC54 was obtained.






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