Difference between revisions of "Team:Aix-Marseille/Amelioration part:BBa K1445000"
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It is immediately apparent from this table that with the BBa_K1445000 biobrick the only antibiotic resistance we recover in phage and PLP is kanamycin resistance carried by M13KO7. | It is immediately apparent from this table that with the BBa_K1445000 biobrick the only antibiotic resistance we recover in phage and PLP is kanamycin resistance carried by M13KO7. | ||
| − | That means the phagemid, which carried | + | That means the phagemid, which carried chloramphanicol resistance, was not packaged at all. |
With the pBlueScript as a control, we can say, due to the BBa_K1445000, we couldn't give chloramphenicol resistance to our infected bacteria. | 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 23:11, 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 |
It is immediately apparent from this table that with the BBa_K1445000 biobrick the only antibiotic resistance we recover in phage and PLP is kanamycin resistance carried by M13KO7. That means the phagemid, which carried chloramphanicol resistance, was not packaged at all. With the pBlueScript as a control, we can say, due to the BBa_K1445000, we couldn't give chloramphenicol resistance to our infected bacteria.
