We had two different plasmids. One plasmid with p5 and p6 on it and one plasmid with phi 29 DNAP and phi 29 TP. We wanted to put this on one plasmid through Gibson Assembly
Results:
There were no plates with colonies, therefore Gibson Assembly has not work.
May
June
July
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Brute Force Experiments 1
Goal:
Putting all separate plasmids in and see what survives
As construction of the plasmids did not work, we just wanted to put all the separate constructs in, to see whether something would grow or not. We induced it with no IPTG, 10 µM IPTG and 50 µM IPTG. We put in the following constructs:
To determine which plasmids were toxic, we put transformed the plasmids separately in BL21 pLysS and set an overnight measurement to measure OD600 to see if the cells were alive.
p01 (control)
p02
p03
p04
Results:
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Characterization of Level 1's
Goal:
Add characterization for the parts page and see if the different promoters strength are visible
To test whether orthogonal replication works in vivo, we transformed all the plasmid with the components needed in pLysS together with a final construct, which is either OriL-Kan-OriR or OriL-GFP-Kan-OriR. This was done with different IPTG concentrations.