Team:Calgary/3 June 2009

From 2009.igem.org

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CTV Interviews + Gel Electrophoresis for Confirmation
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WIKI CODING HERE
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* Was interviewed by CTV news team regarding iGEM and how engineers can contribute to this team
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* Ran a 1% gel to confirm whether pSB1AC3 and pSB1AK3 are the appropriate size. We did several different types of enzyme digestion on the previous day.
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Exploration Results : <br>
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Simbiology assumes that all reactions are elementary reactions. For stochastic simulation only the Chemical kinetic and Hill kinetic equations work. Simbiology also produces a diagram of all the reactions input . The diagram view is helpful in establishing the connections between species when they react or form.
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Exploration Results :
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The Simbiology simulation graphs produced have a generic y-axis value of state which signifies concentration .  
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Simbiology assumes that all reactions are elementary reactions. For stochastic simulation only the Chemical kinetic and Hill kinetic equations work. Simbiology also produces a diagram of all the reactions input . The diagram view is helpfull in establishing the connections between species when they react or form.
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The simbiology simulation graphs produced have a generic y-axis value of state which signifies concentration .  
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Started to look at other Toolboxes given with the Matlab package.  
Started to look at other Toolboxes given with the Matlab package.  
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Visualization of Previous Restriction Digest
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WIKI CODING HERE
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*Visualization of psB1AC3, psB1AK3 and LuxOD47E in pCR.2.1-TOPO vector restriction digest
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*Added 10X Orange dye and ran on a 1% agarose gel, expected band sizes: psB1AC3/psB1AK3 ~ 3.0 KB,
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*Lane 1- Ladder
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Lanes 2-6 psB1AC3
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Lanes 7-11 psB1AK3
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Lane 8- LuxOU
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Lane 9-Lux PQ
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Lane 10- Lux CDABE
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Lane 11- LuxOD47E
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Lane 12- LuxOD47A
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*Analysis: From the gel we can conclude that LuxPQ and LuxCDABE were not cut with EcoRI as we do not see the expected band sizes in these lanes.  The others look good.  We will proceed by sending LuxPQ in TOPO vector and LuxCDABE in TOPO vector for DNA sequencing.
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*Today we also had a vistt from CTV News, a news station in Calgary.  We showed them around the labs an they took some footage of our lab team in action!
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Marketing and Outreach for June 3rd 2009
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WIKI CODING HERE
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Today, our iGEM Calgary team was interviewed by the CTV news courtesy of Shelly Makrugin. In her story she covered areas such as what iGEM is, what is synthetic biology, what is our project this year and what could be the potential applications of our project. The most important thing that excited her was our competition against teams such as Harvard and Stanford.
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IMAN
 
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Since I was leading our iGEM Calgary team at the Campus Fair, I started making preparations for the campus fair.
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WIKI CODING HERE
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JAMIE
 
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JEREMY
 
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WIKI CODING HERE
 
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Working with the Listen Event
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WIKI CODING HERE
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Everything has to be added into chat so answers to questions are entered into the chat window, which I do not really like since mistakes are easy to make and I also keep communicating with a black board that uses one of the channels I am using as well.
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Learned about the listen event:
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* Uses channels and messages and keeps track of a avatar’s id so this will be very useful for: Giving out inventory, dividing messages up
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Cycle of the PCR are complete now and an amplified piece of DNA, which is rezzed beside it and can be taken into the avatar’s inventory to be used for something else later on like running a gel.
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Began looking into biobrick cloning techniques. Reviewed how the parts are put together. If A part is the recepient and B part is the donor, and when B part needs to be put behind of A, the recepient A is cut with SpeI and PstI sites and the donor is cut with XbaI and PstI. When B needs to be put in front of A, the recepient A is cut with EcoRI and Xba, and the donor B is cut with EcoRI and SpeI. When cut Xba and Spe sites meet, it creates a scar, making the construction permenant. (It won't cut apart with the enzymes we use)
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Began looking into biobrick cloning techniques. Reviewed how the parts are put together. If A part is the recipient and B part is the donor, and when B part needs to be put behind of A, the recipient A is cut with SpeI and PstI sites and the donor is cut with XbaI and PstI. When B needs to be put in front of A, the recipient A is cut with EcoRI and Xba, and the donor B is cut with EcoRI and SpeI. When cut Xba and Spe sites meet, it creates a scar, making the construction permanent. (It won't cut apart with the enzymes we use)
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MANDY
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PRIMA
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CTV interviews
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WIKI CODING HERE
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Today, I continued to follow up with companies to whom I had sent the sponsorship package. I made a couple cold calls to companies who didn't provide an email address on their websites. First I tried to get a hold of the person in-charge of marketing/sponsorship, tell them a little about the project, what iGEM is and why we're looking for sponsors. I spoke to a few company representatives and most of them were very excited about the project. I asked for their email address and sent the sponsorship package to them. I'll follow up with them next week.
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VICKI
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Gel of the vector verification performed yesterday
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WIKI CODING HERE
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Purpose: to visualise the results of yesterday’s vector verification
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Expected restriction digest output:
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Biobrick digest (2 fragments)
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* psB1AC3/psB1AK3: ~3.0 kb
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* ccdB (p1010): ~675 bp
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TOPO digest (2 fragments)
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* TOPO (T/A and Blunt): ~3.5 kb
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* LuxOU: ~2.0 kb
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* LuxPQ: ~4.0 kb
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* LuxCDABE: ~6.0 kb
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PRIMA
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* LuxOD47E: ~1.4 kb
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* LuxOD47A: ~1.4 kb
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Results:
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Gel and lane description is included below. LuxPQ did not work, so it will need to be sequenced.
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Sequencing of LuxPQ in TOPO Blunt II
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WIKI CODING HERE
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Purpose: As mentioned above, the results for LuxPQ are very questionable. Since they cannot be explained with restriction digests, hopefully sequencing will alleviate some of the confusion.
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Materials and methods:
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We inserted 7.3 uL of LuxPQ in TOPO into a mini PCR tube so that around 730 ng of DNA were present for sequencing. We used the sp6 forward primer and the T7 reverse primer.
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Results:
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Much like our previous sequencing attempts, these were questionable. The electronic copy will be inserted shortly.
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Latest revision as of 03:57, 19 October 2009

University of Calgary

UNIVERSITY OF CALGARY



THIS MONTH

June
MTWTFSS
[http://2009.igem.org/Team:Calgary/1_June_2009 1] [http://2009.igem.org/Team:Calgary/2_June_2009 2] [http://2009.igem.org/Team:Calgary/3_June_2009 3] [http://2009.igem.org/Team:Calgary/4_June_2009 4] [http://2009.igem.org/Team:Calgary/5_June_2009 5] [http://2009.igem.org/Team:Calgary/6_June_2009 6] [http://2009.igem.org/Team:Calgary/7_June_2009 7]
[http://2009.igem.org/Team:Calgary/8_June_2009 8] [http://2009.igem.org/Team:Calgary/9_June_2009 9] [http://2009.igem.org/Team:Calgary/10_June_2009 10] [http://2009.igem.org/Team:Calgary/11_June_2009 11] [http://2009.igem.org/Team:Calgary/12_June_2009 12] [http://2009.igem.org/Team:Calgary/13_June_2009 13] [http://2009.igem.org/Team:Calgary/14_June_2009 14]
[http://2009.igem.org/Team:Calgary/15_June_2009 15] [http://2009.igem.org/Team:Calgary/16_June_2009 16] [http://2009.igem.org/Team:Calgary/17_June_2009 17] [http://2009.igem.org/Team:Calgary/18_June_2009 18] [http://2009.igem.org/Team:Calgary/19_June_2009 19] [http://2009.igem.org/Team:Calgary/20_June_2009 20] [http://2009.igem.org/Team:Calgary/21_June_2009 21]
[http://2009.igem.org/Team:Calgary/22_June_2009 22] [http://2009.igem.org/Team:Calgary/23_June_2009 23] [http://2009.igem.org/Team:Calgary/24_June_2009 24] [http://2009.igem.org/Team:Calgary/25_June_2009 25] [http://2009.igem.org/Team:Calgary/26_June_2009 26] [http://2009.igem.org/Team:Calgary/27_June_2009 27] [http://2009.igem.org/Team:Calgary/28_June_2009 28]
[http://2009.igem.org/Team:Calgary/29_June_2009 29] [http://2009.igem.org/Team:Calgary/30_June_2009 30]


NOTEBOOK PAGE INDEX
Carol
Chinmoyee
Emily
Fahd
Iman
Jamie
Jeremy
Katie
Kevin
Mandy
Patrick
Prima
Stefan
Vicki



CALENDAR

May
MTWTFSS
        [http://2009.igem.org/Team:Calgary/1_May_2009 1] [http://2009.igem.org/Team:Calgary/2_May_2009 2] [http://2009.igem.org/Team:Calgary/3_May_2009 3]
[http://2009.igem.org/Team:Calgary/4_May_2009 4] [http://2009.igem.org/Team:Calgary/5_May_2009 5] [http://2009.igem.org/Team:Calgary/6_May_2009 6] [http://2009.igem.org/Team:Calgary/7_May_2009 7] [http://2009.igem.org/Team:Calgary/8_May_2009 8] [http://2009.igem.org/Team:Calgary/9_May_2009 9] [http://2009.igem.org/Team:Calgary/10_May_2009 10]
[http://2009.igem.org/Team:Calgary/11_May_2009 11] [http://2009.igem.org/Team:Calgary/12_May_2009 12] [http://2009.igem.org/Team:Calgary/13_May_2009 13] [http://2009.igem.org/Team:Calgary/14_May_2009 14] [http://2009.igem.org/Team:Calgary/15_May_2009 15] [http://2009.igem.org/Team:Calgary/16_May_2009 16] [http://2009.igem.org/Team:Calgary/17_May_2009 17]
[http://2009.igem.org/Team:Calgary/18_May_2009 18] [http://2009.igem.org/Team:Calgary/19_May_2009 19] [http://2009.igem.org/Team:Calgary/20_May_2009 20] [http://2009.igem.org/Team:Calgary/21_May_2009 21] [http://2009.igem.org/Team:Calgary/22_May_2009 22] [http://2009.igem.org/Team:Calgary/23_May_2009 23] [http://2009.igem.org/Team:Calgary/24_May_2009 24]
[http://2009.igem.org/Team:Calgary/25_May_2009 25] [http://2009.igem.org/Team:Calgary/26_May_2009 26] [http://2009.igem.org/Team:Calgary/27_May_2009 27] [http://2009.igem.org/Team:Calgary/28_May_2009 28] [http://2009.igem.org/Team:Calgary/29_May_2009 29] [http://2009.igem.org/Team:Calgary/30_May_2009 30] [http://2009.igem.org/Team:Calgary/31_May_2009 31]


June
MTWTFSS
[http://2009.igem.org/Team:Calgary/1_June_2009 1] [http://2009.igem.org/Team:Calgary/2_June_2009 2] [http://2009.igem.org/Team:Calgary/3_June_2009 3] [http://2009.igem.org/Team:Calgary/4_June_2009 4] [http://2009.igem.org/Team:Calgary/5_June_2009 5] [http://2009.igem.org/Team:Calgary/6_June_2009 6] [http://2009.igem.org/Team:Calgary/7_June_2009 7]
[http://2009.igem.org/Team:Calgary/8_June_2009 8] [http://2009.igem.org/Team:Calgary/9_June_2009 9] [http://2009.igem.org/Team:Calgary/10_June_2009 10] [http://2009.igem.org/Team:Calgary/11_June_2009 11] [http://2009.igem.org/Team:Calgary/12_June_2009 12] [http://2009.igem.org/Team:Calgary/13_June_2009 13] [http://2009.igem.org/Team:Calgary/14_June_2009 14]
[http://2009.igem.org/Team:Calgary/15_June_2009 15] [http://2009.igem.org/Team:Calgary/16_June_2009 16] [http://2009.igem.org/Team:Calgary/17_June_2009 17] [http://2009.igem.org/Team:Calgary/18_June_2009 18] [http://2009.igem.org/Team:Calgary/19_June_2009 19] [http://2009.igem.org/Team:Calgary/20_June_2009 20] [http://2009.igem.org/Team:Calgary/21_June_2009 21]
[http://2009.igem.org/Team:Calgary/22_June_2009 22] [http://2009.igem.org/Team:Calgary/23_June_2009 23] [http://2009.igem.org/Team:Calgary/24_June_2009 24] [http://2009.igem.org/Team:Calgary/25_June_2009 25] [http://2009.igem.org/Team:Calgary/26_June_2009 26] [http://2009.igem.org/Team:Calgary/27_June_2009 27] [http://2009.igem.org/Team:Calgary/28_June_2009 28]
[http://2009.igem.org/Team:Calgary/29_June_2009 29] [http://2009.igem.org/Team:Calgary/30_June_2009 30]


July
MTWTFSS
    [http://2009.igem.org/Team:Calgary/1_July_2009 1] [http://2009.igem.org/Team:Calgary/2_July_2009 2] [http://2009.igem.org/Team:Calgary/3_July_2009 3] [http://2009.igem.org/Team:Calgary/4_July_2009 4] [http://2009.igem.org/Team:Calgary/5_July_2009 5]
[http://2009.igem.org/Team:Calgary/6_July_2009 6] [http://2009.igem.org/Team:Calgary/7_July_2009 7] [http://2009.igem.org/Team:Calgary/8_July_2009 8] [http://2009.igem.org/Team:Calgary/9_July_2009 9] [http://2009.igem.org/Team:Calgary/10_July_2009 10] [http://2009.igem.org/Team:Calgary/11_July_2009 11] [http://2009.igem.org/Team:Calgary/12_July_2009 12]
[http://2009.igem.org/Team:Calgary/13_July_2009 13] [http://2009.igem.org/Team:Calgary/14_July_2009 14] [http://2009.igem.org/Team:Calgary/15_July_2009 15] [http://2009.igem.org/Team:Calgary/16_July_2009 16] [http://2009.igem.org/Team:Calgary/17_July_2009 17] [http://2009.igem.org/Team:Calgary/18_July_2009 18] [http://2009.igem.org/Team:Calgary/19_July_2009 19]
[http://2009.igem.org/Team:Calgary/20_July_2009 20] [http://2009.igem.org/Team:Calgary/21_July_2009 21] [http://2009.igem.org/Team:Calgary/22_July_2009 22] [http://2009.igem.org/Team:Calgary/23_July_2009 23] [http://2009.igem.org/Team:Calgary/24_July_2009 24] [http://2009.igem.org/Team:Calgary/25_July_2009 25] [http://2009.igem.org/Team:Calgary/26_July_2009 26]
[http://2009.igem.org/Team:Calgary/27_July_2009 27] [http://2009.igem.org/Team:Calgary/28_July_2009 28] [http://2009.igem.org/Team:Calgary/29_July_2009 29] [http://2009.igem.org/Team:Calgary/30_July_2009 30] [http://2009.igem.org/Team:Calgary/31_July_2009 31]


August
MTWTFSS
          [http://2009.igem.org/Team:Calgary/1_August_2009 1] [http://2009.igem.org/Team:Calgary/2_August_2009 2]
[http://2009.igem.org/Team:Calgary/3_August_2009 3] [http://2009.igem.org/Team:Calgary/4_August_2009 4] [http://2009.igem.org/Team:Calgary/5_August_2009 5] [http://2009.igem.org/Team:Calgary/6_August_2009 6] [http://2009.igem.org/Team:Calgary/7_August_2009 7] [http://2009.igem.org/Team:Calgary/8_August_2009 8] [http://2009.igem.org/Team:Calgary/9_August_2009 9]
[http://2009.igem.org/Team:Calgary/10_August_2009 10] [http://2009.igem.org/Team:Calgary/11_August_2009 11] [http://2009.igem.org/Team:Calgary/12_August_2009 12] [http://2009.igem.org/Team:Calgary/13_August_2009 13] [http://2009.igem.org/Team:Calgary/14_August_2009 14] [http://2009.igem.org/Team:Calgary/15_August_2009 15] [http://2009.igem.org/Team:Calgary/16_August_2009 16]
[http://2009.igem.org/Team:Calgary/17_August_2009 17] [http://2009.igem.org/Team:Calgary/18_August_2009 18] [http://2009.igem.org/Team:Calgary/19_August_2009 19] [http://2009.igem.org/Team:Calgary/20_August_2009 20] [http://2009.igem.org/Team:Calgary/21_August_2009 21] [http://2009.igem.org/Team:Calgary/22_August_2009 22] [http://2009.igem.org/Team:Calgary/23_August_2009 23]
[http://2009.igem.org/Team:Calgary/24_August_2009 24] [http://2009.igem.org/Team:Calgary/25_August_2009 25] [http://2009.igem.org/Team:Calgary/26_August_2009 26] [http://2009.igem.org/Team:Calgary/27_August_2009 27] [http://2009.igem.org/Team:Calgary/28_August_2009 28] [http://2009.igem.org/Team:Calgary/29_August_2009 29] [http://2009.igem.org/Team:Calgary/30_August_2009 30]
[http://2009.igem.org/Team:Calgary/31_August_2009 31]


September
MTWTFSS
  [http://2009.igem.org/Team:Calgary/1_September_2009 1] [http://2009.igem.org/Team:Calgary/2_September_2009 2] [http://2009.igem.org/Team:Calgary/3_September_2009 3] [http://2009.igem.org/Team:Calgary/4_September_2009 4] [http://2009.igem.org/Team:Calgary/5_September_2009 5] [http://2009.igem.org/Team:Calgary/6_September_2009 6]
[http://2009.igem.org/Team:Calgary/7_September_2009 7] [http://2009.igem.org/Team:Calgary/8_September_2009 8] [http://2009.igem.org/Team:Calgary/9_September_2009 9] [http://2009.igem.org/Team:Calgary/10_September_2009 10] [http://2009.igem.org/Team:Calgary/11_September_2009 11] [http://2009.igem.org/Team:Calgary/12_September_2009 12] [http://2009.igem.org/Team:Calgary/13_September_2009 13]
[http://2009.igem.org/Team:Calgary/14_September_2009 14] [http://2009.igem.org/Team:Calgary/15_September_2009 15] [http://2009.igem.org/Team:Calgary/16_September_2009 16] [http://2009.igem.org/Team:Calgary/17_September_2009 17] [http://2009.igem.org/Team:Calgary/18_September_2009 18] [http://2009.igem.org/Team:Calgary/19_September_2009 19] [http://2009.igem.org/Team:Calgary/20_September_2009 20]
[http://2009.igem.org/Team:Calgary/21_September_2009 21] [http://2009.igem.org/Team:Calgary/22_September_2009 22] [http://2009.igem.org/Team:Calgary/23_September_2009 23] [http://2009.igem.org/Team:Calgary/24_September_2009 24] [http://2009.igem.org/Team:Calgary/25_September_2009 25] [http://2009.igem.org/Team:Calgary/26_September_2009 26] [http://2009.igem.org/Team:Calgary/27_September_2009 27]
[http://2009.igem.org/Team:Calgary/28_September_2009 28] [http://2009.igem.org/Team:Calgary/29_September_2009 29] [http://2009.igem.org/Team:Calgary/30_September_2009 30]


October
MTWTFSS
      [http://2009.igem.org/Team:Calgary/1_October_2009 1] [http://2009.igem.org/Team:Calgary/2_October_2009 2] [http://2009.igem.org/Team:Calgary/3_October_2009 3] [http://2009.igem.org/Team:Calgary/4_October_2009 4]
[http://2009.igem.org/Team:Calgary/5_October_2009 5] [http://2009.igem.org/Team:Calgary/6_October_2009 6] [http://2009.igem.org/Team:Calgary/7_October_2009 7] [http://2009.igem.org/Team:Calgary/8_October_2009 8] [http://2009.igem.org/Team:Calgary/9_October_2009 9] [http://2009.igem.org/Team:Calgary/10_October_2009 10] [http://2009.igem.org/Team:Calgary/11_October_2009 11]
[http://2009.igem.org/Team:Calgary/12_October_2009 12] [http://2009.igem.org/Team:Calgary/13_October_2009 13] [http://2009.igem.org/Team:Calgary/14_October_2009 14] [http://2009.igem.org/Team:Calgary/15_October_2009 15] [http://2009.igem.org/Team:Calgary/16_October_2009 16] [http://2009.igem.org/Team:Calgary/17_October_2009 17] [http://2009.igem.org/Team:Calgary/18_October_2009 18]
[http://2009.igem.org/wiki/index.php?title=Team:Calgary/19_October_2009&preload=Team:Calgary/NotebookPreload&action=edit 19] [http://2009.igem.org/wiki/index.php?title=Team:Calgary/20_October_2009&preload=Team:Calgary/NotebookPreload&action=edit 20] [http://2009.igem.org/wiki/index.php?title=Team:Calgary/21_October_2009&preload=Team:Calgary/NotebookPreload&action=edit 21] [http://2009.igem.org/wiki/index.php?title=Team:Calgary/22_October_2009&preload=Team:Calgary/NotebookPreload&action=edit 22] [http://2009.igem.org/wiki/index.php?title=Team:Calgary/23_October_2009&preload=Team:Calgary/NotebookPreload&action=edit 23] [http://2009.igem.org/wiki/index.php?title=Team:Calgary/24_October_2009&preload=Team:Calgary/NotebookPreload&action=edit 24] [http://2009.igem.org/wiki/index.php?title=Team:Calgary/25_October_2009&preload=Team:Calgary/NotebookPreload&action=edit 25]
[http://2009.igem.org/wiki/index.php?title=Team:Calgary/26_October_2009&preload=Team:Calgary/NotebookPreload&action=edit 26] [http://2009.igem.org/wiki/index.php?title=Team:Calgary/27_October_2009&preload=Team:Calgary/NotebookPreload&action=edit 27] [http://2009.igem.org/wiki/index.php?title=Team:Calgary/28_October_2009&preload=Team:Calgary/NotebookPreload&action=edit 28] [http://2009.igem.org/wiki/index.php?title=Team:Calgary/29_October_2009&preload=Team:Calgary/NotebookPreload&action=edit 29] [http://2009.igem.org/wiki/index.php?title=Team:Calgary/30_October_2009&preload=Team:Calgary/NotebookPreload&action=edit 30] [http://2009.igem.org/wiki/index.php?title=Team:Calgary/31_October_2009&preload=Team:Calgary/NotebookPreload&action=edit 31]



JUNE 3, 2009


CAROL

CTV Interviews + Gel Electrophoresis for Confirmation

  • Was interviewed by CTV news team regarding iGEM and how engineers can contribute to this team
  • Ran a 1% gel to confirm whether pSB1AC3 and pSB1AK3 are the appropriate size. We did several different types of enzyme digestion on the previous day.


CHINMOYEE

More Simbiology

Exploration Results :
Simbiology assumes that all reactions are elementary reactions. For stochastic simulation only the Chemical kinetic and Hill kinetic equations work. Simbiology also produces a diagram of all the reactions input . The diagram view is helpful in establishing the connections between species when they react or form.

The Simbiology simulation graphs produced have a generic y-axis value of state which signifies concentration .

Started to look at other Toolboxes given with the Matlab package.


EMILY

Visualization of Previous Restriction Digest

  • Visualization of psB1AC3, psB1AK3 and LuxOD47E in pCR.2.1-TOPO vector restriction digest
  • Added 10X Orange dye and ran on a 1% agarose gel, expected band sizes: psB1AC3/psB1AK3 ~ 3.0 KB,
  • Lane 1- Ladder


Lanes 2-6 psB1AC3
Lanes 7-11 psB1AK3
Lane 8- LuxOU
Lane 9-Lux PQ
Lane 10- Lux CDABE
Lane 11- LuxOD47E
Lane 12- LuxOD47A

  • Analysis: From the gel we can conclude that LuxPQ and LuxCDABE were not cut with EcoRI as we do not see the expected band sizes in these lanes. The others look good. We will proceed by sending LuxPQ in TOPO vector and LuxCDABE in TOPO vector for DNA sequencing.
  • Today we also had a vistt from CTV News, a news station in Calgary. We showed them around the labs an they took some footage of our lab team in action!


FAHD

Marketing and Outreach for June 3rd 2009

Today, our iGEM Calgary team was interviewed by the CTV news courtesy of Shelly Makrugin. In her story she covered areas such as what iGEM is, what is synthetic biology, what is our project this year and what could be the potential applications of our project. The most important thing that excited her was our competition against teams such as Harvard and Stanford.

Since I was leading our iGEM Calgary team at the Campus Fair, I started making preparations for the campus fair.


KATIE

Working with the Listen Event

Everything has to be added into chat so answers to questions are entered into the chat window, which I do not really like since mistakes are easy to make and I also keep communicating with a black board that uses one of the channels I am using as well.

Learned about the listen event:

  • Uses channels and messages and keeps track of a avatar’s id so this will be very useful for: Giving out inventory, dividing messages up

Cycle of the PCR are complete now and an amplified piece of DNA, which is rezzed beside it and can be taken into the avatar’s inventory to be used for something else later on like running a gel.


KEVIN

Cloning Technique

Began looking into biobrick cloning techniques. Reviewed how the parts are put together. If A part is the recipient and B part is the donor, and when B part needs to be put behind of A, the recipient A is cut with SpeI and PstI sites and the donor is cut with XbaI and PstI. When B needs to be put in front of A, the recipient A is cut with EcoRI and Xba, and the donor B is cut with EcoRI and SpeI. When cut Xba and Spe sites meet, it creates a scar, making the construction permanent. (It won't cut apart with the enzymes we use)


PRIMA

CTV interviews

Today, I continued to follow up with companies to whom I had sent the sponsorship package. I made a couple cold calls to companies who didn't provide an email address on their websites. First I tried to get a hold of the person in-charge of marketing/sponsorship, tell them a little about the project, what iGEM is and why we're looking for sponsors. I spoke to a few company representatives and most of them were very excited about the project. I asked for their email address and sent the sponsorship package to them. I'll follow up with them next week.


VICKI

Gel of the vector verification performed yesterday

Purpose: to visualise the results of yesterday’s vector verification

Expected restriction digest output:

Biobrick digest (2 fragments)

  • psB1AC3/psB1AK3: ~3.0 kb
  • ccdB (p1010): ~675 bp

TOPO digest (2 fragments)

  • TOPO (T/A and Blunt): ~3.5 kb
  • LuxOU: ~2.0 kb
  • LuxPQ: ~4.0 kb
  • LuxCDABE: ~6.0 kb
  • LuxOD47E: ~1.4 kb
  • LuxOD47A: ~1.4 kb

Results:

Gel and lane description is included below. LuxPQ did not work, so it will need to be sequenced.


Sequencing of LuxPQ in TOPO Blunt II

Purpose: As mentioned above, the results for LuxPQ are very questionable. Since they cannot be explained with restriction digests, hopefully sequencing will alleviate some of the confusion.

Materials and methods: We inserted 7.3 uL of LuxPQ in TOPO into a mini PCR tube so that around 730 ng of DNA were present for sequencing. We used the sp6 forward primer and the T7 reverse primer.

Results: Much like our previous sequencing attempts, these were questionable. The electronic copy will be inserted shortly.

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