Team:Calgary/26 June 2009

From 2009.igem.org

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Descriptive Title of What You're Doing
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Constructing LuxPQ into psB1AC3 plasmid
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WIKI CODING HERE
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Purpose: to insert linear LuxPQ into a BBK vector in order to continue the construction of signaling circuit with the BBK standard. Linear LuxPQ with BBK restriction sites was digested with the following sets of enzymes: XbaI and PstI, EcoRI and PstI. Using the same sets of enzymes, the psB1AC3 vector was also digested for 2 hours at 37ºC. Subsequent phosphotase and ligase treatment followed, as well as transformation into TOP10 competent cells. These cells were then plated on LB+ chlor plates overnight at 37ºC.
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Revision as of 20:22, 3 September 2009

University of Calgary

UNIVERSITY OF CALGARY



THIS MONTH

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June 26, 2009


CAROL

Descriptive Title of What You’re Doing

WIKI CODING HERE

CHINMOYEE

Looking into More Papers

Stochastic Models of Gene Expression
Stochastic mechanisms in gene expression
A stochastic model of Escherichia coli AI-2 quorum signal circuit reveals alternative synthesis pathways --- has stochastic rate constants


EMILY

Descriptive Title of What You're Doing

WIKI CODING HERE


FAHD

Team presentations

Today we had our team presentations. I presented on behalf of the Human Practices aspect of our project


IMAN

Graphs, Rules, and First Steps in Division

Graph section of our model is completed this week. As it was mentioned, now we have two general types of graphs, one showing the change of concentration of an element in all the cells, and the other one shows the change of concentration of an element in each cell. There is a menu designed in result section that users can use to choose which one of these types of graphs they would like to observe. This week, I also had a meeting with the other modeling team and we compared our models together. As a result of that meeting, we had to change and edit some of our evolution rules. Changing a rule in this model could not be done easily. When a rule changes, many functions should change too. For instance, some functions read rules and calculate the possibilities of their applications. Other functions apply the rules to the environment by eliminating the reactants and adding products. Therefore, I spent some time this week to edit the existing rules and adding the new rules asked by Thane and the equation based modeling group. Another important aspect that was discussed in the meeting was “DIVISION”. I have started to implement this function to our model. This function should get an environment as input and applies division to the environment and pass the new environment with double number of cells as its output. I had also two other meetings this week. Lindsay meeting and iGEM meeting.


JAMIE

Descriptive Title of What You're Doing

WIKI CODING HERE


JEREMY

Constructing LuxPQ into psB1AC3 plasmid

Purpose: to insert linear LuxPQ into a BBK vector in order to continue the construction of signaling circuit with the BBK standard. Linear LuxPQ with BBK restriction sites was digested with the following sets of enzymes: XbaI and PstI, EcoRI and PstI. Using the same sets of enzymes, the psB1AC3 vector was also digested for 2 hours at 37ºC. Subsequent phosphotase and ligase treatment followed, as well as transformation into TOP10 competent cells. These cells were then plated on LB+ chlor plates overnight at 37ºC.


KATIE

Putting Scripts into the Lab Equipment

I wrote out the pseudo code for the water bath script and the donor and recipient tubes for restriction digest this week as well as well as completing the first basic working version inside of test tubes instead of a basic primitive that were initially going to be given to the avatar so they could move and work with them anywhere they want, but know I believe that the tubes will remain in the lab and only be moved when they have to be sent to the water bath, heating block, etc. The order script is not complete yet, but that is because I am still changing it since other items will still have to be added to give something back to avatars.

Presently the test tubes will:

  • Tell you if the item you added is correct or not. The tubes also tell you it is correct to delete something too, but hopefully other avatars will not have this permission. I will need to remove items that are not needed as well, which is where the cleaning script comes in as well as getting rid of items once they are no longer needed
  • When touched, the tube will ask if you would like the circuit part to go behind or in front of another and then will search for specific enzymes depending on your choice and tells me if i can continue or not

I will continue to expand o this and hopefully incorporate phosphatase treatment into the activity in the upcoming week.


KEVIN

Descriptive of What You're Doing

WIKI CODING HERE


MANDY

Descriptive Title of What You're Doing

WIKI CODING HERE


PATRICK

Descriptive Title of What You're Doing

WIKI CODING HERE


PRIMA

Continuation of Marketing

I shadowed Carol while she did restriction digest. She was doing construction. Her vector was apsB1AK3. However, since I didn't go into the lab very often, I wasn't sure what she was digesting. I just watched the processes and she just explained the restriction sites.


STEFAN

Helping out in the lab

I've been finishing some objects that are going to be pit in the laboratory this week. Most notable are the microscope, various test tubes and scripted water bath. At the same time, arrangement adjustments we're carried out, creating walls for the labs, and an entry area as well as perfecting the Key Pad door I have been working on.

Calgary Microscope 001.png



VICKI

Colony PCR of J13002+LuxOD47A and LuxOD47A+B0015 bacteria

Purpose: Cultures grew successfully on plates last night. This will help confirm which of those colonies took up the plasmids that we wanted to, and that the plasmids that were taken up include properly-constructed genes in the right order.

Protocol: The colony PCR was conducted in accordance with that outlined on the protocol page. Platinum Taq polymerase was used, along with BBk construction primers. 5 colonies were tested for each construct, along with one negative control (for a total of 11 tubes being amplified).

Results: The gel is included below. The negative control results are peculiar, but are not raising any concern as the contamination band is not found anywhere else on the gel. Some of the LuxOD47A stock was run to serve as a size comparison. From this, it appears that the LuxOD47A+B0015 parts were successfully constructed and transformed, along with colonies 2 and 4 of the J13002+LuxOD47A parts.



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