M13 part improvement
As we use a phage like particle, we needed to use the M13 phagemid. 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.
After that, we executed the Large quantities of helper phage protocol and infected GM1 bacteria with the resulting phage like particle.
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 |