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Virginia/21 July 2008
From 2008.igem.org
Where We Stand
- Genetic Attenuator
- Measurement Plasmid
- Activating Promoter with Arabinos Xincomplete
- Suspicion that measurement plasmid may not work in DB3.1 cells we are cutting out cell death gene and growing up in DH5alpha cells
- Ligation
- RFP fluorescence finished
- GFP fluorescence XIncomplete
- need Green to fluoresce on its own before we ligate all necessary parts for the testing plasmid
- need to get both Red and Green Fluorescence working on one plasmid either through the arabinose inudced easurement plasmid or by ligating each of the genes together but we are having problems with both methods
- BioPlastic
- BP1 gene
- Status: PRET
- Gene is available in previous form for assembly as an operon
- need to perform SDS-PAGE on PRET form to verify phaA is being produced by the gene
- BP2 gene
- Status: RE
- Need to Ligate inducible Promoter
- Need to Ligate Terminator
- Need to perform SDS-PAGE
- BP3 gene
- Status: RE
- Need to Ligate inducible Promoter
- Need to Ligate Terminator
- Need to Perform SDS-PAGE
- After all 3 are functioning independently
- Transform cells with all 3 genes on seperate vectors
- Create a single Operon with all 3 genes included to transform cells
- Perform SDS-PAGE
- Perform GCMS
- Adding to iGEM library
- Order Fluorescent proteins Finished
- Order AntiBiotic Resistances -------
- Order Construction Sites Incomplete
- Submit Bioplastic genes to registry Incomplete
Overnight Results
- Overnight Broths of PRET BP1, RE BP2, RE BP3
- NONE of the plated ligations grew
Goals
- Research whats needed for SDS-PAGE proceadure
- Update Protocol Page
- Update Wike Calender
- Research Terminators not included in the registry
- Model - Read Kinetic Eq. paper and make some progress on model
- Gel of Digested DNA
- Ligate promoter to A12(RE BP2) and A13(RE BP3)
- Ligate PRE RFP to new RET GFP
