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Team:UC Berkeley/ProjectOverview
From 2008.igem.org
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# Prepro Parts | # Prepro Parts | ||
We made parts of the format {promoter}{rbs}{prepro}{phoA}{term}. The promoter we used was {pBad}, and we tried 3 different ribosome binding sites with each prepro. We plated the bacteria we transformed on two plates, both with p-nitrophenyl phosphate, but one had arabinose and one did not. We screened successful rbs/prepro combinations by looking for combinations which caused blue colonies on the plate with arabinose and white colonies on the plate without arabinose. This could also be done in liquid culture. In preparation for later composite parts, we made sure that the {rbs}{prepro} part was an intermediate when making our construction tree. | We made parts of the format {promoter}{rbs}{prepro}{phoA}{term}. The promoter we used was {pBad}, and we tried 3 different ribosome binding sites with each prepro. We plated the bacteria we transformed on two plates, both with p-nitrophenyl phosphate, but one had arabinose and one did not. We screened successful rbs/prepro combinations by looking for combinations which caused blue colonies on the plate with arabinose and white colonies on the plate without arabinose. This could also be done in liquid culture. In preparation for later composite parts, we made sure that the {rbs}{prepro} part was an intermediate when making our construction tree. | ||
| + | |||
| + | # Promoters | ||
| + | Make parts of the format {promoter}{rbs}{GFP}{term}, where we want to make the last three, {rbs}{GFP}{term}, into one part and test the different OD dependent promoters: hns, spv, bolA, ftsAZ, ftsQ, rrnB P1, and Ptet (as a positive control, which we already have). As a negative control, we will have no promoter. <br> | ||
| + | |||
| + | The experiment involves diluting saturated cultures and growing them at 37 C, take out at the different time points and test the fluorescence to determine at what OD they start to turn on. <br> | ||
| + | |||
| + | '''Sound-dependent Promoter''' <br> | ||
| + | Make parts of the format {Psound}{rbs}{GFP}{term}. Grow in culture and apply sound for 30 min. Then measure fluorescence. | ||
| + | |||
| + | # Amplifier | ||
| + | Make parts of the format {Pbad}{spuR}{Pspu2}{rbs}{GFP}{term}, where {spuR}{Pspu2} is a composite part, and {rbs}{GFP}{term} is a composite part. For the control, we will have {Pbad}{rbs}{GFP}{term}. We will grow the culture to mid-log, induce w/0.2x arabinose and measure the fluorescence after 1 hour. | ||
| + | |||
| + | |||
Revision as of 22:01, 17 June 2008
Contents |
| You can write a background of your team here. Give us a background of your team, the members, etc. Or tell us more about something of your choosing. |  |
|
Tell us more about your project. Give us background. Use this is the abstract of your project. Be descriptive but concise (1-2 paragraphs) |  Your team picture |
| Team Example 2 |
| Home | The Team | The Project | Parts Submitted to the Registry | Modeling | Notebook |
|---|
(Or you can choose different headings. But you must have a team page, a project page, and a notebook page.)
Overall project
Our project is centered around a system that will allow sound-induced lysis o E. coli.
Project Details
Part 2
The Experiments
Testing Individual Composite Parts
- Prepro Parts
We made parts of the format {promoter}{rbs}{prepro}{phoA}{term}. The promoter we used was {pBad}, and we tried 3 different ribosome binding sites with each prepro. We plated the bacteria we transformed on two plates, both with p-nitrophenyl phosphate, but one had arabinose and one did not. We screened successful rbs/prepro combinations by looking for combinations which caused blue colonies on the plate with arabinose and white colonies on the plate without arabinose. This could also be done in liquid culture. In preparation for later composite parts, we made sure that the {rbs}{prepro} part was an intermediate when making our construction tree.
- Promoters
Make parts of the format {promoter}{rbs}{GFP}{term}, where we want to make the last three, {rbs}{GFP}{term}, into one part and test the different OD dependent promoters: hns, spv, bolA, ftsAZ, ftsQ, rrnB P1, and Ptet (as a positive control, which we already have). As a negative control, we will have no promoter.
The experiment involves diluting saturated cultures and growing them at 37 C, take out at the different time points and test the fluorescence to determine at what OD they start to turn on.
Sound-dependent Promoter
Make parts of the format {Psound}{rbs}{GFP}{term}. Grow in culture and apply sound for 30 min. Then measure fluorescence.
- Amplifier
Make parts of the format {Pbad}{spuR}{Pspu2}{rbs}{GFP}{term}, where {spuR}{Pspu2} is a composite part, and {rbs}{GFP}{term} is a composite part. For the control, we will have {Pbad}{rbs}{GFP}{term}. We will grow the culture to mid-log, induce w/0.2x arabinose and measure the fluorescence after 1 hour.
