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

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===Applications of BBa_K320004===
 
===Applications of BBa_K320004===
  
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We transformed E.Coli and Asaia with this biobrick in the pSBC3 backbone plasmid vector. We plated those cultur  respectively on Lb+Agar+Kanamycin and Gly+Agar+Kanamycin.  
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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''.  
  
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We observe that on both plates we obtain colonnies and that in negativ controls (E.Coli and Asaia without BBaK320004 in pSBC3 backbone vector) we obtain non colonnies.
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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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Put Picture of the plates
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We transfected both ''E. coli'' and ''Asaia'' with the plasmid.
  
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In conclusion we assume that our resitance to Kanamycin works and that the origin in replication also works in Asaia.
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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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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===
 
===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

UNIQe68838f9b0717803-partinfo-00000000-QINU UNIQe68838f9b0717803-partinfo-00000001-QINU