Team:Calgary/2 June 2009

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<div class="heading">NOTEBOOK PAGE INDEX</div>
 
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<a href="#Carol">Carol</a><br>
 
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<a href="#Chinmoyee">Chinmoyee</a><br>
 
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<a href="#Emily">Emily</a><br>
 
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<a href="#Fahd">Fahd</a><br>
 
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<a href="#Iman">Iman</a><br>
 
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<a href="#Jamie">Jamie</a><br>
 
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<a href="#Jeremy">Jeremy</a><br>
 
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<a href="#Katie">Katie</a><br>
 
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<a href="#Kevin">Kevin</a><br>
 
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<a href="#Mandy">Mandy</a><br>
 
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<a href="#Patrick">Patrick</a><br>
 
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<a href="#Prima">Prima</a><br>
 
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<a href="#Stefan">Stefan</a><br>
 
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<a href="#Vicki">Vicki</a><br>
 
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<div class="heading">CALENDAR</div>
 
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* Used GenElute Plasmid Miniprep Kit (Sigma-Aldrich), used as according to the manufacturers instructions.
* Used GenElute Plasmid Miniprep Kit (Sigma-Aldrich), used as according to the manufacturers instructions.
*Results:
*Results:
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* Set up restriction enzyme digest to ensure that biobrick sites are in the vector. The following are the reactions that were done:
* Set up restriction enzyme digest to ensure that biobrick sites are in the vector. The following are the reactions that were done:
Reaction 1: pSB1AC3 - cut with NotI
Reaction 1: pSB1AC3 - cut with NotI
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CHINMOYEE
 
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Descriptive Title of What You're Doing
 
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WIKI CODING HERE
 
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Purification of psB1AC3 & psB1AK3 vectors
Purification of psB1AC3 & psB1AK3 vectors
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*Protocol: Used GenElute Plasmid Miniprep Kit (Sigma-Aldrich), lot # 048k6062, used as according to the manufacturers instructions.
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*Protocol: Used GenElute Plasmid Miniprep Kit (Sigma-Aldrich), lot # 048k6062, used as according to the manufacturer's instructions.
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*Results:
*Results:
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FAHD
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Restriction digest of psB1AC3 and psB1AK3 vectors as well as LuxOD47E in pCR.2.1-TOPO vector
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WIKI CODING HERE
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*Objective: To verify the presence of restriction sites in the two BBK vectors as well as in our BBK gene in the pCR.2.1-TOPO vector
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*Protocol:
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5 reactions for psB1AC3 vector, 5 reactions for psB1AK3 vector, 1 reaction for gene of interest (LuxOD47E in pCR2.1-TOPO vector).
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BBK Vector Reaction List
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Reaction 1- Not1 + React 3 Buffer
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Reaction 2- EcoRI + SpeI + React Buffer 4
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Reaction 3- XbaI + PstI + React 2 Buffer
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Reaction 4- EcoRI + PstI + React 2 Buffer
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Reaction 5- XbaI + SpeI + React 4 Buffer
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BBK vector tube preparation
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8 uL Plasmid (psB1AC3 or psB1AK3)
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2 uL appropriate React Buffer
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1 uL of each appropriate restrcition enzyme
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ddH20 up to 20 uL
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TOPO vector tube preparation
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10 uL DNA (LuxOD47E in pCR2.1- TOPO vector)
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7 uL ddH20
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2 uL React 3 Buffer
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1 uL NotI restriction enzymes
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Left to digest overnight in waterbath at 37°C.
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IMAN
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FAHD
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Descriptive Title of What You're Doing
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Marketing and Media for June 2nd 2009
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WIKI CODING HERE
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Today, I started filling out funding applications for some pharmaceutical companies. Some of these companies include Pfizer Canada, Merck Frosst Canada and Glaxosmithkline Canada. I did not get a chance to complete them but I am still working on it.
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I also set up a TV media appointment with the NUTV news for June the 16th 2009.
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Biobrick Vector Isolation and Verification + Sponsorship Package work
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Today the iGEM Lab team worked on isolating psB1AC3 and psB1AK3.  Digestion with all enzymes in the prefix and suffix was used to confirm the presence of the restriction sites.<br /><br />
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WIKI CODING HERE
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I also spent some time drafting up the sponsorship package for companies.
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Isolation and verification of pSB1AC3 and pSB1AK3
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WIKI CODING HERE
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The iGEM lab team members worked together today to help each other perfect the method of plasmid isolation. Plasmids psB1AC3 and psB1AK3 were isolated using Sigma GenElute Plasmid Mini-prep kit, and the concentration and purity of plasmid was measured using the NanoDrop Spectrophotometer. See Emily’s notebook for today (above) for results.
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In order to verify that these vectors only had the death gene ccdB (675bp) in them, RD was set up with combinations of the following restriction enzymes: EcoRI, XbaI, PstI, NotI and SpeI. A gel was run with these products, as well as products from a RD set up with various genes of interest in their respective TOPO vectors (all cut with EcoRI).
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Genome Island
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WIKI CODING HERE
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I got a chance to finally explore Genome Island properly and went through some of the activities on the island, which contains informative areas on Mendelian genetics and includes quizzes, slide shows etc. There was a tour of the island, a scavenger hunt, a garden containing plasmids as well as a tower with a variety of lab equipment where Mandy was able to talk to the owner of the island.
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I am hoping we will be able to make our island more interactive than this, but it will depend on how difficult I find the scripting language.
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Plasmid Purification/isolation and verification of pSB1AC3
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WIKI CODING HERE
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<b>Plasmid Isolation</b>
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To isoloate pSB1AC3 plasmid from TOP10 cells, a miniprep on them was done using Sigma's GenElute plasmid Miniprep Kit (sigma). pSB1AC3 plasmids are needed for potential plasmid switches.
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Table 1. Purity and concentration of pSB1AC3 measured with Nanodrop utility and Spectrophotometer
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| '''Plasmid''' || '''260/280'''    || '''260/230''' || '''Concentration [ng/μL]'''
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| pSB1AC3 C3 || 2.05 || 2.42 || 134.4
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| pSB1AC3 C4 || 1.95 || 2.23 || 55.6
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<b>Restriction Digest</b>
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To verify if the biobrick restriction sites are present in pSB1AC3, restrition digest was done using all the different combinations of enzymes:<br>
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NotI, EcorI+SpeI, XbaI+PstI, EcorI+PstI and XbaI+SpeI
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[[Image:Calgary_2009.06.03.BBK_Verification_RD_copy.png|700px]]
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Second Life Teleporter
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WIKI CODING HERE
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The teleportation grid now has buttons that turn black, indicating that the corresponding biobrick sphere is in use, and they remain that way for half an hour. Unfortunately, now that we look back on the spheres, it is dificult to have enough room to get all the parts for biobricking into them, as they can only be 10x10x10 in size as a maximum.
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Today was also our shopping day in preparation for activity materials for the University Campus Fair.
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WIKI CODING HERE
 
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Biotech companies
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WIKI CODING HERE
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Today I made an general email which I could sent to all Biotech companies. I had it checked by the rest of the marketing team. I had tailored some of the emails to target companies who had sponsored the building of the O'Brian labs and donated/offered quotes in the past. Since we had established a friendly long-term relationship with some of these companies, I wrote an email specifically to them regarding sponsoring this year's iGEM project.
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STEFAN
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VICKI
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Verification of vectors
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WIKI CODING HERE
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Purpose: Check the BBk and TOPO vector tubes and confirm that they actually contain what the labels claim they do.
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Materials and methods:
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1. The plasmids (BBk and ccdB) came in E. coli. We performed a mini-prep to isolate them from the bacteria, in accordance with our plasmid isolation procedure. We were dealing with both psB1AC3 and psB1AK3 plasmids.
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2. We evaluated the DNA concentrations with the Nanodrop spectrophotometer
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3. We confirmed the contents with a restriction digest.
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Restriction digest volumes
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For TOPO plasmids: take 10 uL LuxPQ, 7 uL ddHOH
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For restriction digest: 1 uL of each restriction enzyme, 2 uL of the relevant buffer, 8 uL of the plasmid and 9 uL of ddHOH – for a total volume of 20 uL
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VICKI
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Reactions (enzymes that were used in cutting and relevant buffers):
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* Tube 1 – NotI (REact 3)
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* Tube 2 – EcoRI and SpeI (REact 4)
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Descriptive Title of What You're Doing
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* Tube 3 – XbaI and PstI (REact 2)
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* Tube 4 – EcoRI and PstI (REact 2)
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* Tube 5 – XbaI and SpeI (REact 4)
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* Tube 6 – TOPO with EcoRI (REact 3)
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WIKI CODING HERE
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Results:
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The restriction digest products were visualised on an agarose gel the following day.
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Latest revision as of 03:53, 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 2, 2009


CAROL

Isolation of Plasmids (pSB1AC3 and pSB1AK3)for Cloning

  • Isolated plasmids (pSB1AK3 and pSB1AC3) from overnight cultures prepared by Thane.
  • Used GenElute Plasmid Miniprep Kit (Sigma-Aldrich), used as according to the manufacturers instructions.
  • Results:
Plasmid 260/280 260/230 Concentration [ng/μL]
psB1AC3 1 1.96 3.48 76.9
psB1AC3 2 1.93 2.29 98.1
psB1AC3 3 1.87 1.88 48.5
psB1AC3 4 1.90 2.11 118.9
psB1AK3 1 2.05 2.42 134.4
psB1AK3 2 1.95 2.23 55.6
psB1AK3 3 1.91 2.22 97.9
psB1AK3 4 2.03 1.92 44.5


  • Set up restriction enzyme digest to ensure that biobrick sites are in the vector. The following are the reactions that were done:

Reaction 1: pSB1AC3 - cut with NotI Reaction 2: pSB1AC3 - cut with EcoRI and SpeI Reaction 3: pSB1AC3 - cut with XbaI and PstI Reaction 4: pSB1AC3 - cut with XbaI and SpeI Reaction 5: pSB1AK3 - cut with NotI Reaction 6: pSB1AK3 - cut with EcorI and SpeI Reaction 7: pSB1AK3 - cut with XbaI and PstI Reaction 8: pSB1AK3 - cut with XbaI and SpeI

  • Incubate reactions at 37oC


EMILY

Purification of psB1AC3 & psB1AK3 vectors

  • Protocol: Used GenElute Plasmid Miniprep Kit (Sigma-Aldrich), lot # 048k6062, used as according to the manufacturer's instructions.


  • Results:
Plasmid 260/280 260/230 Concentration [ng/μL]
psB1AC3 1 1.96 3.48 76.9
psB1AC3 2 1.93 2.29 98.1
psB1AC3 3 1.87 1.88 48.5
psB1AC3 4 1.90 2.11 118.9
psB1AK3 1 2.05 2.42 134.4
psB1AK3 2 1.95 2.23 55.6
psB1AK3 3 1.91 2.22 97.9
psB1AK3 4 2.03 1.92 44.5

Restriction digest of psB1AC3 and psB1AK3 vectors as well as LuxOD47E in pCR.2.1-TOPO vector

  • Objective: To verify the presence of restriction sites in the two BBK vectors as well as in our BBK gene in the pCR.2.1-TOPO vector


  • Protocol:

5 reactions for psB1AC3 vector, 5 reactions for psB1AK3 vector, 1 reaction for gene of interest (LuxOD47E in pCR2.1-TOPO vector).

BBK Vector Reaction List

Reaction 1- Not1 + React 3 Buffer
Reaction 2- EcoRI + SpeI + React Buffer 4
Reaction 3- XbaI + PstI + React 2 Buffer
Reaction 4- EcoRI + PstI + React 2 Buffer
Reaction 5- XbaI + SpeI + React 4 Buffer

BBK vector tube preparation
8 uL Plasmid (psB1AC3 or psB1AK3)
2 uL appropriate React Buffer
1 uL of each appropriate restrcition enzyme
ddH20 up to 20 uL

TOPO vector tube preparation
10 uL DNA (LuxOD47E in pCR2.1- TOPO vector)
7 uL ddH20
2 uL React 3 Buffer
1 uL NotI restriction enzymes

Left to digest overnight in waterbath at 37°C.


FAHD

Marketing and Media for June 2nd 2009

Today, I started filling out funding applications for some pharmaceutical companies. Some of these companies include Pfizer Canada, Merck Frosst Canada and Glaxosmithkline Canada. I did not get a chance to complete them but I am still working on it.

I also set up a TV media appointment with the NUTV news for June the 16th 2009.


JAMIE

Biobrick Vector Isolation and Verification + Sponsorship Package work

Today the iGEM Lab team worked on isolating psB1AC3 and psB1AK3. Digestion with all enzymes in the prefix and suffix was used to confirm the presence of the restriction sites.

I also spent some time drafting up the sponsorship package for companies.


JEREMY

Isolation and verification of pSB1AC3 and pSB1AK3

The iGEM lab team members worked together today to help each other perfect the method of plasmid isolation. Plasmids psB1AC3 and psB1AK3 were isolated using Sigma GenElute Plasmid Mini-prep kit, and the concentration and purity of plasmid was measured using the NanoDrop Spectrophotometer. See Emily’s notebook for today (above) for results.

In order to verify that these vectors only had the death gene ccdB (675bp) in them, RD was set up with combinations of the following restriction enzymes: EcoRI, XbaI, PstI, NotI and SpeI. A gel was run with these products, as well as products from a RD set up with various genes of interest in their respective TOPO vectors (all cut with EcoRI).


KATIE

Genome Island

I got a chance to finally explore Genome Island properly and went through some of the activities on the island, which contains informative areas on Mendelian genetics and includes quizzes, slide shows etc. There was a tour of the island, a scavenger hunt, a garden containing plasmids as well as a tower with a variety of lab equipment where Mandy was able to talk to the owner of the island.

I am hoping we will be able to make our island more interactive than this, but it will depend on how difficult I find the scripting language.


KEVIN

Plasmid Purification/isolation and verification of pSB1AC3

Plasmid Isolation To isoloate pSB1AC3 plasmid from TOP10 cells, a miniprep on them was done using Sigma's GenElute plasmid Miniprep Kit (sigma). pSB1AC3 plasmids are needed for potential plasmid switches.

Table 1. Purity and concentration of pSB1AC3 measured with Nanodrop utility and Spectrophotometer

Plasmid 260/280 260/230 Concentration [ng/μL]
pSB1AC3 C3 2.05 2.42 134.4
pSB1AC3 C4 1.95 2.23 55.6



Restriction Digest To verify if the biobrick restriction sites are present in pSB1AC3, restrition digest was done using all the different combinations of enzymes:
NotI, EcorI+SpeI, XbaI+PstI, EcorI+PstI and XbaI+SpeI
Calgary 2009.06.03.BBK Verification RD copy.png


MANDY

Second Life Teleporter

The teleportation grid now has buttons that turn black, indicating that the corresponding biobrick sphere is in use, and they remain that way for half an hour. Unfortunately, now that we look back on the spheres, it is dificult to have enough room to get all the parts for biobricking into them, as they can only be 10x10x10 in size as a maximum. Today was also our shopping day in preparation for activity materials for the University Campus Fair.


PRIMA

Biotech companies

Today I made an general email which I could sent to all Biotech companies. I had it checked by the rest of the marketing team. I had tailored some of the emails to target companies who had sponsored the building of the O'Brian labs and donated/offered quotes in the past. Since we had established a friendly long-term relationship with some of these companies, I wrote an email specifically to them regarding sponsoring this year's iGEM project.


VICKI

Verification of vectors

Purpose: Check the BBk and TOPO vector tubes and confirm that they actually contain what the labels claim they do.

Materials and methods: 1. The plasmids (BBk and ccdB) came in E. coli. We performed a mini-prep to isolate them from the bacteria, in accordance with our plasmid isolation procedure. We were dealing with both psB1AC3 and psB1AK3 plasmids.

2. We evaluated the DNA concentrations with the Nanodrop spectrophotometer

3. We confirmed the contents with a restriction digest.

Restriction digest volumes

For TOPO plasmids: take 10 uL LuxPQ, 7 uL ddHOH

For restriction digest: 1 uL of each restriction enzyme, 2 uL of the relevant buffer, 8 uL of the plasmid and 9 uL of ddHOH – for a total volume of 20 uL

Reactions (enzymes that were used in cutting and relevant buffers):

  • Tube 1 – NotI (REact 3)
  • Tube 2 – EcoRI and SpeI (REact 4)
  • Tube 3 – XbaI and PstI (REact 2)
  • Tube 4 – EcoRI and PstI (REact 2)
  • Tube 5 – XbaI and SpeI (REact 4)
  • Tube 6 – TOPO with EcoRI (REact 3)

Results:

The restriction digest products were visualised on an agarose gel the following day.

Retrieved from "http://2009.igem.org/Team:Calgary/2_June_2009"