Difference between revisions of "Part:BBa K320004:Experience"

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===Applications of BBa_K320004===
 
===Applications of BBa_K320004===
  
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This BioBrick has two main fonction, it makes the bacteria to resistant to the Antibiotic Kanamycin and it provides an origin of replication working for ''E.Coli'' and ''Asaia''.  
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This BioBrick has two main functions, it makes the bacteria to resistant to the Antibiotic Kanamycin and it provides an origin of replication working for ''E. coli'' and ''Asaia''.  
  
 
To test the BioBrick, we added it in the pSBC3 vector. This plasmid provides a resistance to Chloramphenicole and an origin of replication which works only for ''E.Coli''.
 
To test the BioBrick, we added it in the pSBC3 vector. This plasmid provides a resistance to Chloramphenicole and an origin of replication which works only for ''E.Coli''.
  
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We transfected both ''E.Coli'' and ''Asaia'' with the plasmid.  
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We transfected both ''E. coli'' and ''Asaia'' with the plasmid.  
  
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The transfected ''E.Coli'' were grown on a plate of LB with Agar containing Kanamycin with a concentration of . This experiment allows us to confirm that our part provides a resistance to Kanamycin.
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The transfected ''E. coli'' were grown on a plate of LB with Agar containing Kanamycin. This experiment confirmed that the part provides the bacteria with resistance to Kanamycin.
  
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The transfected ''Asaia'' were grown on a plate of Gly with Agar containing Kanamycin with a concentration of ...(according to our [http://2010.igem.org/Team:EPF_Lausanne/Project_asaia "characterisation of Asaia"]). This experiment allows us to confirm that the origin of replication and that ''Asaia'' aquired resistance to Kanamycin.
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Until the wiki freeze we haven't been able to successfully transform Asaia with this construct. We don't know if that is caused by either the origin, the resistance gene or the fact that the plasmid contains two origins at the same time (our positive control [http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1885625/?tool=pubmed pHM2-GFP plasmid], which contains the same origin but a non-biobrick kanamycin resistance, works).
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We also made two negative controls with E.Coli or Asaia without plamsid.
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We observed that there was no colonies on the negative control plates, in opposition to the presence of colonies for our transfected bacteria.
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Put Picture of the plates
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In conclusion we assume that our resitance to Kanamycin works and that the origin in replication works for ''Asaia''.
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===User Reviews===
 
===User Reviews===

Latest revision as of 22:34, 27 October 2010

This experience page is provided so that any user may enter their experience using this part.
Please enter how you used this part and how it worked out.

Applications of BBa_K320004

This BioBrick has two main functions, it makes the bacteria to resistant to the Antibiotic Kanamycin and it provides an origin of replication working for E. coli and Asaia.

To test the BioBrick, we added it in the pSBC3 vector. This plasmid provides a resistance to Chloramphenicole and an origin of replication which works only for E.Coli.

We transfected both E. coli and Asaia with the plasmid.

The transfected E. coli were grown on a plate of LB with Agar containing Kanamycin. This experiment confirmed that the part provides the bacteria with resistance to Kanamycin.

Until the wiki freeze we haven't been able to successfully transform Asaia with this construct. We don't know if that is caused by either the origin, the resistance gene or the fact that the plasmid contains two origins at the same time (our positive control [http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1885625/?tool=pubmed pHM2-GFP plasmid], which contains the same origin but a non-biobrick kanamycin resistance, works).

User Reviews

UNIQ44f8f6c40977361e-partinfo-00000000-QINU UNIQ44f8f6c40977361e-partinfo-00000001-QINU