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- Because the light sensor pieces are not cutting efficiently, we are working on digesting RBS in a more effective method
1. Digest RBS w/ buffer 3 & Pst
2. Acetate precipitate
3. Digest RBS w/ buffer 2 & Spe
- We are doing this because PST is only 75% active in buffer 2 but is only 100% active in buffer 3
- Proceeded to digest parts: B0034 (3) (Spe), I15008 (Xba & Pst), I15009 (Xba & Pst), I13453 (Spe & Pst), B0015 (Eco & Xba)
- And the bands showed quite well
- This led to the successful ligation of RBS w/ I15008, RBS w/ I15009, and I15010 with B0015
- Ligations for the ON switch and the initiator continue to fail several more times, but more knowledge and insight is gained. What is the cause for the failed ligations? Perhaps it is the low concentration of the gel purification product? We will nanodrop our miniprep and every step of the way to figure out where the concentration is being lost. Are pieces not being cut? We noticed a bunch of colonies on our vector controls. Are we doing something wrong in the experiment? We will run some tests next week.
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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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