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

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|10⁻²
 
|10⁻²
 
|0
 
|0
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|386
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|186
 
|0
 
|0
 
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|10⁻³
 
|10⁻³
 
|0
 
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−
With this spreadsheet, we can see that pBlueScript
+
With this spreadsheet, we can see that, with the BBa_K1445000 biobrick, the only antibiotic resistance we can have is the kanamycin. That means the phagemid packaging didn't work. 
 +
With the pBlueScript as a control, we can say, due to the BBa_K1445000, we couldn't give chloramphenicol resistance to our infected bacteria.

Revision as of 22:28, 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 overnight growth, we counted the colonies. Here are the results for the M13 from the BBa_K1445000, and the pBlueScript II plasmid that we used (all measurements were made in triplicate):

pBlueScript II BBa_K1445000
Ampicilin Kanamycin Ampicilin + Kanalycin Chloramphenicol Kanamycin Chloramphenicol + Kanamycin
1µL 162 606 1 1µL 0 957 0
10⁻² 1 33 0 10⁻² 0 186 0
10⁻³ 0 1 0 10⁻³ 0 28 0
500µL uncountable uncountable 7 500µL 0 uncountable 0


With this spreadsheet, we can see that, with the BBa_K1445000 biobrick, the only antibiotic resistance we can have is the kanamycin. That means the phagemid packaging didn't work. With the pBlueScript as a control, we can say, due to the BBa_K1445000, we couldn't give chloramphenicol resistance to our infected bacteria.