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The transformation from the day before worked. There were no colonies is the insert and the competent cells control as expected. There were colonies in the backbones control and in the gibson transformation plates.
1) Colony PCR
we used one Fw primer we designed, annealing inside the INP sequence and the RV primer from iGEM registry.
Tm = 61°C using Phusion polymerase and 1min30sec extension time.
- 8 colonies from G.A. IYS plate
- 8 colonies from G.A. ISY plate
- 2 colonies respectively from IYS and ISY backbone controls plates
- 2 colonies from the BBa_K523013 miniprep template transformation plate

2) Colonies inoculation for microscopy
- Inoculation in 3ml LB + Chloramphenicol + IPTG of the 16 gibson assembly products and the 2 BBa_K523013 miniprep colonies.
- Inoculation in 3ml LB of normal competent cells to be used as negative controls