3-8-11
Overnight cultures of PBAD-cI-LVA-DT, PBAD-LasR-LVA-DT, PBAD-RBS-GFP-DT, PBAD-pSB1C3 grew.
Plasmid prep of the samples. Measuring with ND1000- Nanodrop showed that all samples contained between 50 and 80 ng/μl
DNA. Samples were prepared and sent for sequencing. Also glycerol stocks had been made of the overnight cultures.
Ligation mixture PBAD-RBS-GFP-DT was used again for transformation.
Transformation was done as usual:
Add to 40μl competent cells E.coli DH5alpha 10μl ligation mixture.
Incubate on ice for 30 min.
Put the cells in a waterbath set on 42 degrees and incubate for 45s.
After the heat shock, place tubes on ice for two minutes.
After the two minutes of incubation, add 1ml of LB medium+25mM Glucose.
Incubate the cells for 1h or 1,5h
Plate cells out on plates with the required antibiotic
Incubate the plates overnight at 37 degrees.
Also, PhybB-RBS-GFP-DT and control strain pGFP-rrnB will be grown in M9 minimal medium for tomorrow's measurements with the
flow cytometer.
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