Team:Aix-Marseille/Amelioration part:BBa K1445000
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):
| 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 |
