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CAROL
Transformation of R0040 (promoter) +pCS26 (vector) into XL Gold cells
- From the overnight digest of R0040 with XhoI and BamHI, the promoter was ligated with vector, pCS26.
- These plasmids were transformed into XL Gold cells and was plated on Kanamycin plates overnight.
Team meeting:
- Lab:discussed about promoter library and the failure to see the expression of luciferase. Also, discussed about the primer design of R0040, so that we can clone the constitutively ON promoter in front of luciferase. This will show us what the luciferase expression will look like.
- Modelling: Discussed about Robustness. The three things that we decided to look at for robustness is: 1. PQ expression (the selection of the 'best' sigma 70 promoter) 2. Temperature Alterations (to see if changes in temperature (from 37oC to another temperature would affect expression) 3. Type of media (LB vs. LM)
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CHINMOYEE
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EMILY
Sequencing of J13002-LuxOD47E-B0015 Construct in psB1AC3
- Sent Colony 3 of my J13002-LuxOD47E-B0015 construct down for sequencing, results will come in on Monday hopefully.
- Explored Secondlife and met with Rob from UChicago.
- Prepared the Human Practices project write-up for the Wiki.
- Team Meeting- Talked about the exploration of seminars in Second Life and what ideas that has given us. Talked about the pending completion of the J13002-LuxOD47E-B0015 curcuit, results will come in on Monday.
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FAHD
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IMAN
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JAMIE
Verification of cl lambda inverter in psB2K3
- Plasmid miniprep of overnight cultures with Sigma Genelute Miniprep kit. Standard manufacturer protocol used; elution in 40µL of ddH2O.
- Vacufuge to concentrate.
- Verification digest with XbaI and PstI. Control: cl lambda part from the Registry distribution plate.
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JEREMY
Isolation of cl lambda; Sequencing for LuxPQ-LuxOU construct
Six overnight cultures of cl lambda colony 1 were prepared with LB-Kanamycin broth. Plasmid was then isolated using the Sigma GenElute MiniPrep kit. Two plasmids were not successfully isolated, but the other four tubes were pooled into two tubes and vacufuged to increase their concentration. A restriction digest with XbaI and PstI was then set up for 2 hours at 37ºC.
The LuxPQ-B-R-LuxOU-B construct in psB1AC3 was sent for sequencing today based on verification techniques performed over the past few days. Primers used include BBK CP F/R and R0040 Reverse.
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KATIE
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KEVIN
cPCR of the reporter circuit
In order to verify cPCR, I have once again ran a cPCR of the reporter circuit, Pqrr4+B0034(RBS)+K082003(GFP+LVA) with Pqrr4 Forward and Biobrick CP Reverse primers. Of course, pTaq was used to amplify the DNA. The product was then ran on 2% agarose gel at 90V. ----the gel is being run right now, will edit---
Distribution of our baking sale posters
Fahd left a stack of Baking sale posters for us to post throughout the building, and they were posted before 11am today.
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MANDY
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PATRICK
Automagic Biobricks
I spent most of today being very productive with the Biobricker portion of the Biobrick Simulator. This is a little user interface upgrade that will permit you to build any biobrick device you please, from a small library of parts implemented in Second Life. In SL terms, the UI is an object owned by the user's avatar, that can be attached to your display. The simulator's UI will have quite a few features to make your digital bioengineering easier, of which the Biobricker is an important one.
I finished working on all of the controls for the biobricker, including some scroll buttons for the parts in the device under construction, and the ability to add parts from the UI's inventory, and delete them too. The Biobricker is all done except for one last control... the build button! This single button will be as much work as the rest of the Biobricker put together, though. The task is to create a copy of each individual Biobrick in world, inform each one where it belongs in the completed device, and link them all together. This will involve a lot of communication back and forth between the user interface object and the biobrick objects out in the world.
The Biobricker is in a very primitive state right now, with letters standing in for where all the pretty icons will be in the final version. So instead here's a screenshot of the Main Menu, with the link to the Biobricker visible.
The UI fits in at the top left corner of the screen. The red letters are buttons, that cause the UI to be redrawn with a new menu for whichever module you've chosen. I hope this gives you an idea of how the UI fits into the overall simulator.
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PRIMA
Continuation of last day's work
Today, I edited the high school presentation proposition again and sent it out to Mrs. Ellechuk. I filled in the lab notebook and filled out the daily updates on the wiki.
I finished typing out the acknowledgement for each mentor/advisor of iGEM Calgary. I also helped to write references for Sonja. Then I finished off with mailing out more newsletters. We also had a very productive team meeting today.
On Monday I have to follow up with the companies which I have been contacting for the past few days. I will also call our hotel in Boston to change a few rooms around.
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STEFAN
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VICKI
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