Team:UC Davis/Notebook

From 2009.igem.org

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July August September Results

July:
        Before beginning the wet lab portion of our project, we have read numerous articles and journals relevant to our proposed project, and have utilized this information to guide our part designs. After significant research, design and redesign, we have settled on the final design for our two key parts, and plan for their construction.
        We have begun ordering the necessary reagents for our work. This includes, selecting the parts required for our project from those provided by the iGEM parts distribution, oligonucleotides for extracting them, and for the synthesis of novel parts via Polymerase Chain Reaction, PCR, and the DNA synthesis of several novel parts from GENEART.
        We have recently started the synthesis and assembly of the initial elements required for the secretion system.

August:
After reading the numerous articles we dug up in regards to our project, we have designed our project; in blue print form of structural wet lab experimentation, as well as the techniques, materials, equipment, and time required to complete our project.
The parts and a majority of the reagents we have ordered necessary for our work have arrived, and we are in the stages of conducting experiments to construct our parts. We have to construct, test, and prove through sequencing as well as other miscellaneous techniques that we truly have the parts we want in our constructs. So the progress for the month of August varies greatly on almost a daily basis.

Apart from the wet lab portion of our project, we have also begun the initial stages of building our wiki. We have begun to revise our information regarding the team, biomedical engineering, and how it is UC Davis’s inauguration year into the iGEM competition.

September:

    We are putting together the final pieces of our intended construct and have designed the tiny details of our experiments’ proper controls.The design itself is useless without making it as a physical part, so we are already building these simultaneously with our experimental constructs.
    The wiki is now up on the iGEM website and is being reviewed countless times to ensure accuracy and appeal.
    We have also designed team t-shirts, which are a tradition of iGEM, and are currently in the process of making them into print.We are also making the presentation power point for presentation at the competition. We are also designed the poster for presentation at the competition.

Results:

Data collection strategy:

Strain:
1. (no induction) ------> Spin cells to seperate cells from media ------> Check for fluorescence or western blot with 6-His Tag
2. (induced with IPTG) ------> Spin cells to seperate cells from media ------> Check for fluorescence or western blot with 6-His Tag

Whole cells were lyced and ran on this gel. The difference in protein concentrations are shown between the different plasmid containing cells. Each well was loaded with a cell containing the vectors labeled on top of the well.

INPNC_SS_GFP
whole cell fluorescence spectrum

Ex: 395 nm
Green = INPNC.SS.GFP induced
red = INPNC.SS.GFP uninduced