Results

Physic

Manage to make a blue LED that emitted light in the correct wavelength
Proved that there was no fluorescens with green and red light (but with blue).
Was not able to measure the light emitted from GFP with a CCD camera.
At some point had some red light emitted from the GFP
Tested multiple set ups (none that proved lasing, at least with our equipment)
Verified that you do not need monochromatic light to get fluorescens

Parts in submission vector:

Cyc1: Successful transformation of E.coli TOP10 with Gibson solution indicated by bacterial growth on LB plates containing chloramphenicol(25 mg/mL). (plate picture)

Results from PCR further confirm an insert of correct length. (Gel picture)

Sequencing results show a successful insertion into pSB1C3. (picture of sequins results)

Nmt1: Successful transformation of E.coli TOP10 with Gibson solution indicated by bacterial growth on LB plates containing chloramphenicol(25 mg/mL). However there were red colonies persent, meaning that they contained an empty vector. (reference to the iGEM page Athanasios found) (plate picture)

Gel from PCR suggested that white colonies do not contain insert, but rather are bacteria that are not chloramphenicol resistant. This occurs when plates are incubated to long and antibiotica has deteriorated. Reason for longer incubation time than recommended is due to the antibiotics used. Chloramphenicol is tougher on bacteria cells than ampicillin and bacteria may need more time to grow. (gel bilde)

Several attempts have given same result. It been theorized that the reason why nmt1 is harder to insert is caused by its DNA secondary structure. (Eric told me, need to find source).

However, this stepback has caused us to modify gibson assembly protocol. Increase incubation to 1h and dilute the gibson solution to remove some molecules in the Gibens Mix causing trouble during transformation. We have diluted 1:3, but increased the volume used in transformation by 3 times. Leading to the same volume of plasmid, but ⅓ of the gibson mix. Results show an increased number of transformed colonies from the diluted solution. (picture plate)

Further results for PCR indicat a insert of right size, followed by sequencing confirmation. (gel picture and sequencing results).

Composite part : (the same as above, if we manage to do it)

INTERLAB 96-Well Transformation Protocol: http://parts.igem.org/Help:Protocols/Transformation#96-Well_Transformation_Protocol Plate reader protocol: https://static.igem.org/mediawiki/2017/8/85/InterLab_2017_Plate_Reader_Protocol.pdf We used both clear and black bottom plates We did not do PCR to conform; we thought the presence of colonies on the plates was sufficient