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

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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.
 
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 concetration of 100 ng/microl ([http://2010.igem.org/Team:EPF_Lausanne/Project_asaia "characterisation of Asaia"]). This experiment confirmed that the origin of replication worked in ''Asaia'' and that the BioBrick conferred resistance to kanamycine.
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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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Of course we also made two negative controls with E. coli or Asaia without plasmid, for which we observed no colonies on any plate.
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We conclude that the resistance to Kanamycin and the origin of replication work for both ''E. coli'' and ''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

UNIQa3fca3ecaa36baf6-partinfo-00000000-QINU UNIQa3fca3ecaa36baf6-partinfo-00000001-QINU