Difference between revisions of "Team:Aix-Marseille/Amelioration part:BBa K1445000"

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{{Aix-Marseille|title=M13 part improvement|toc=__NOTOC__}}
 
{{Aix-Marseille|title=M13 part improvement|toc=__NOTOC__}}
  
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As we use a phage like particle, we needed to use the [[Team:Aix-Marseille/M13_Design#Phagemid|M13 phagemid]].
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As we wanted to make phage like particles, we needed to use a [[Team:Aix-Marseille/M13_Design#Phagemid|M13 phagemid]].
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Results weren’t good, after a transformation using TG1 bacteria with the [http://parts.igem.org/Part:BBa_K1445000 BBa_K1445000], we picked up the colonies on a chloramphenicol plate and infected them with our M13KO7 phage like particle.  
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Initially we tried using [http://parts.igem.org/Part:BBa_K1445000 BBa_K1445000].
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After a transforming TG1 with the biobrick we selected colonies, on chloramphenicol, and infected them with helper phage (M13KO7).  
  
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After that, we executed the [[Team:Aix-Marseille/Experiments/Protocols#Production_of_large_quantities_of_helper_phage|Large quantities of helper phage]] protocol and infected GM1 bacteria with the resulting phage like particle.
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We then purified [[Team:Aix-Marseille/Experiments/Protocols#Production_of_large_quantities_of_helper_phage|phage]] and infected ''E.coli'' GM1 bacteria with the resulting preparation.
  
 
After an overnight grow, we counted the colonies.
 
After an overnight grow, we counted the colonies.

Revision as of 21:58, 1 November 2017

M13 part improvement

As we wanted to make phage like particles, we needed to use a M13 phagemid. Initially we tried using [http://parts.igem.org/Part:BBa_K1445000 BBa_K1445000]. After a transforming TG1 with the biobrick we selected colonies, on chloramphenicol, and infected them with helper phage (M13KO7).

We then purified phage and infected E.coli GM1 bacteria with the resulting preparation.

After an overnight grow, we counted the colonies. Here are the results for the M13 from the BBa_K1445000, and the pBlueScript II plasmid that we used:


In this experiment, for better results, everything was done three times, and these results are averages.

BBa_K1445000 pBlueScript II
Ampicilin Kanamycin Ampicilin + Kanalycin Chloramphenicol Kanamycin Chloramphenicol + Kanamycin
1µL 162 606 1 1µL 0 957 0
10⁻² 1 33 0 10⁻² 0 386 0
10⁻³ 0 1 0 10⁻³ 0 58 0
500µL Cell sheet Cell sheet 7 500µL 0 Cell sheet 0