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Starting Up
- Hydrated parts: BBa_B0034, BBa_C0051, BBa_B0015, BBa_R0082.
- Transformed these parts into DH5α competent cells.
RBS to BBa_C0051
- Ligate the RBS and BBa_C0051 fragments together and transform on carb plates.
- Depending on the outcome of visible colonies on the experimental and control plates, a PCR screening may be needed.
Assembly notes
- It is most practical to cut RBS as a vector because RBS is less than 100 bp long as an insert, making it hard to see and extract on the gel.
- The sticky ends from the XbaI and SpeI, when ligated together, will form an 8 bp scar. Thus, XbaI and SpeI will no longer be able to cut here.
'RBS & BBa_C0051' to Terminator
- Cultured cells that have parts: 'RBS & BBa_C0051', BBa_B0015
- Miniprepped 'RBS & BBa_C0051' and BBa_B0015
- Ligate the fragments together and transform on kan plates.
- Depending on the outcome of visible colonies on the experimental and control plates, a PCR screening may be needed.
'RBS & BBa_C0051 & stop' to BBa_R0082
- Cultured cells that have parts: 'RBS & BBa_C0051 & stop', BBa_R0082
- Miniprepped 'RBS & BBa_C0051& stop' and BBa_R0082
- Ligate the fragments together and transform on carb plates.
- Depending on the outcome of visible colonies on the experimental and control plates, a PCR screening may be needed.
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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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