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- Light sensor: Parts A & B were ligated successfully! Almost no colonies were present on the control plates.
- Samples of C0051 colony #2 were sent in for sequencing to ensure that it is C0051.
- Initiator progress: now that RBS-C0051 is finally ligated together, it’s now time to ligate RBS-C0051 to a double terminator (B0015). Though what we found odd was that after culturing 6 colonies of B0015, only 2 grew. We find this odd because usually all cultures fail or all are successful.
- We nanodropped our B0015 samples: concentrations were low
- We then ran a diagnostic gel…which then showed that we had nothing!
- We then miniprepped the 2 B0015 cultures that grew and then transformed them into new cells. We believe the low concentration is because we tried culturing cells from an old plate. Hence, the cells must have kicked out the B0015 plasmid (with the kanamycin antibiotic resistance) because they don’t need it anymore. This way, we get a “fresh batch” of B0015 plasmids. B0015 previously digested for other parts were obtained from an old miniprep.
- Meanwhile, we have made an interesting discovery:
- RBS-C0051 (which has no promoter) was successfully ligated to RBS-RFP-F1610 (no previous expression)...and it expresses!
- We will send in parts for sequencing.
- We have investigated this phenomena further by redigesting RBS-C0051 (SP) and C0051.
Strangely, RBS-C0051 was shorter than C0051. Maybe the RBS-C0051 is actually RBS-I12006.
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We would like to take a moment to thank all of our sponsors for their very generous donations, as we could not have done this without your help!
We would also like to thank and acknowledge:
Our Advisors
Marc Facciotti
Ilias Tagkopoulos
Technical Guidance
David Larsen
Andrew Yao
Visiting iGEMer
Jia Li of Zhejiang University (TEAM ZJU-China)
cI Promoter Screen
Drew Endy - Stanford
Thomas Schneider - NIH
Want to sponsor us? Send an email to mtfacciotti@ucdavis.edu to discuss various ways you can help! :)
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