Construction and Testing
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Revision as of 13:03, 26 October 2007
Contents |
Construction Tree
The diagram below outlines our parallel construction plan, from individual registry parts to the final half-adder. Because many people were working on the project at once, it is designed to keep everyone organised.
Diagram details:
- Vector resistances are shown, with shades of blue representing ampicillin and shades of red representing kanamycin so it is easy to check which media is needed.
- Fragments all have names small enough to write on a microfuge tube cap. To make the subtle distinction between a coding sequence and a coding sequence and a ribosome binding site, we use a ' on the computer and an underline when writing. A double ' or double underline, indicates a coding sequence with a ribosome binding site and a transcriptional terminator. This helps keep our microfuge caps legible.
- Expected fragment sizes are shown at every node so gel results can be analyzed quickly.
- Arrows show which way DNA is moving and which restriction enzymes to use for cutting. Hollow arrows are present when two vectors of the same resistance are being joined and treatment with alkaline phosphatase or a gel extraction is needed to remove contaminating vector.
- The numbers 1-2-3-3.5-4 show the major steps that need to be done at each step of way. Our protocols are all written around this system.
Testing Constructs
Making use of flurorescent proteins(GFP and RFP) as reporter genes - use of indicuble promoters to...
Comparing basal and induced expression levels for the quorum sensing promoters
Testing functionality of the various promoters
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