Team:Calgary/23 June 2009

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June 23, 2009
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JUNE 23, 2009
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Multiple Miniprep isolations
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WIKI CODING HERE
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* Attempted 15 miniprep reactions on both non-mutated and mutated <i>luxCDABE</i> plasmid.
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* The isolated plasmid was pooled together and attempted to concentrate the plasmid by spinning the tube in a vacifuge. After several hours (2.5 hours) the concentration was higher.
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* Tried ethanol precipitation earlier in the day but there was no precipitate found when done.
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Verification of NotI Digest
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WIKI CODING HERE
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*Objective: To do one last verification that LuxOD47E is in the psB1AC3 vector before we send it down for DNA Sequecning.  We will do a NotI digest with
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*Started with Plasmid Isolation this morning (Sigma).
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*Used 1000 Nanodrop Spectrophotometer to determine concentrations.
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*Set up a restriction digest with NotI enzyme and REact Buffer 3, left to digest in the 37 C waterbath overnight.
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Marketing for 23rd June 2009
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WIKI CODING HERE
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Today, I helped finish the July newsletter which will be ready be next week to sent out to our sponsors. Also I e-mailed some research and development companies and will do a follow-up soon.
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Verify B0015-R0040 and luxOU-B0015 construction
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WIKI CODING HERE
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[[Image:Calgary_luxOU_B0015-R0040+luxOU-B0015_NotI.jpg | 500px]]
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Figure. 1% agarose gel (100V) for NotI verification digest of construction yielding BBa_B0015-BBa_R0040 and luxOU(XbaI or EcoRI)-BBa_B0015.  200ng of plasmid was cut with NotI for 2 hours at 37oC. BBa_B0015-BBa_R0040 C1 is lane 2) and C4 is lane 4.  luxOU(XbaI)-BBa_B0015 C3 is in lane 4 and C4 is in lane 5.  luxOU(EcoRI)-BBa_B0015 C3 is in lane 6 and C4 in lane 7.  Uncut luxOU in psB1AC3 (EcoRI C1 and XbaI C2) were used as positive controls in lanes 8 and 9, respectively.  Expected band sizes for the BBa_B0015-BBa_R0040 construction are 3.2kb (psB1AK3) and 183bp.  Expected band sizes for the luxOU-BBa_B0015 construction are 3kb (psB1AC3) and 2.1kb.  5μL of GeneRuler 1kb Plus DNA Ladder was loaded. 
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BBa_B0015-BBa_R0040 C1 and luxOU(XbaI)-BBa_B0015 C4 were sent for sequencing with BBK-CP-F and BBK-CP-R as sequencing primers. Sequencing results were checked for vector contamination using “VecScreen”, protein sequences using “Blastx” and sequence conformation using “Blast2” (National Center for Biotechnology Information, MD).
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Isolating LuxPQ in psB1AK3 from overnights
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Purpose: isolate LuxPQ in psB1AK3 from overnights of Colony 4 to continue construction of signaling circuit. Plasmid was isolated from colony 4 of luxPQ. This was done using the SIGMA’s Genelute plasmid Miniprep Kit.  Plasmid purity and concentration were measured using the NanoDrop Spectrophotometer.
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To obtain more isolated R0040, B0015, J1302, and Pqrr4, miniprep (plasmid isolation) was done on them.
To obtain more isolated R0040, B0015, J1302, and Pqrr4, miniprep (plasmid isolation) was done on them.
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Blog Making
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WIKI CODING HERE
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Put together a workable blog and other online accounts for our team to update each other on what they've been up to, with an established schedule and everything :) Now I just have to figure out how to get it onto our wiki, or if I have to transfer each post manually from http://igemcalgary.blogspot.com
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Marketing & Sponsorship
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WIKI CODING HERE
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After constantly following up with companies over the past few weeks, I successfully recruited Alberta Research Council to sponsor the University of Calgary's iGEM team. They have generously donated %250 dollars this year.
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We also edited the June newsletter and discussed it with the rest of the marketing team prior to sending it to all the companies we had contacted (including those who have declined our proposal).
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Latest revision as of 06:26, 20 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



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 23, 2009


CAROL

Multiple Miniprep isolations

  • Attempted 15 miniprep reactions on both non-mutated and mutated luxCDABE plasmid.
  • The isolated plasmid was pooled together and attempted to concentrate the plasmid by spinning the tube in a vacifuge. After several hours (2.5 hours) the concentration was higher.
  • Tried ethanol precipitation earlier in the day but there was no precipitate found when done.


EMILY

Verification of NotI Digest

  • Objective: To do one last verification that LuxOD47E is in the psB1AC3 vector before we send it down for DNA Sequecning. We will do a NotI digest with
  • Started with Plasmid Isolation this morning (Sigma).
  • Used 1000 Nanodrop Spectrophotometer to determine concentrations.
  • Set up a restriction digest with NotI enzyme and REact Buffer 3, left to digest in the 37 C waterbath overnight.


FAHD

Marketing for 23rd June 2009

Today, I helped finish the July newsletter which will be ready be next week to sent out to our sponsors. Also I e-mailed some research and development companies and will do a follow-up soon.


JAMIE

Verify B0015-R0040 and luxOU-B0015 construction

Calgary luxOU B0015-R0040+luxOU-B0015 NotI.jpg

Figure. 1% agarose gel (100V) for NotI verification digest of construction yielding BBa_B0015-BBa_R0040 and luxOU(XbaI or EcoRI)-BBa_B0015. 200ng of plasmid was cut with NotI for 2 hours at 37oC. BBa_B0015-BBa_R0040 C1 is lane 2) and C4 is lane 4. luxOU(XbaI)-BBa_B0015 C3 is in lane 4 and C4 is in lane 5. luxOU(EcoRI)-BBa_B0015 C3 is in lane 6 and C4 in lane 7. Uncut luxOU in psB1AC3 (EcoRI C1 and XbaI C2) were used as positive controls in lanes 8 and 9, respectively. Expected band sizes for the BBa_B0015-BBa_R0040 construction are 3.2kb (psB1AK3) and 183bp. Expected band sizes for the luxOU-BBa_B0015 construction are 3kb (psB1AC3) and 2.1kb. 5μL of GeneRuler 1kb Plus DNA Ladder was loaded.

BBa_B0015-BBa_R0040 C1 and luxOU(XbaI)-BBa_B0015 C4 were sent for sequencing with BBK-CP-F and BBK-CP-R as sequencing primers. Sequencing results were checked for vector contamination using “VecScreen”, protein sequences using “Blastx” and sequence conformation using “Blast2” (National Center for Biotechnology Information, MD).


JEREMY

Isolating LuxPQ in psB1AK3 from overnights

Purpose: isolate LuxPQ in psB1AK3 from overnights of Colony 4 to continue construction of signaling circuit. Plasmid was isolated from colony 4 of luxPQ. This was done using the SIGMA’s Genelute plasmid Miniprep Kit. Plasmid purity and concentration were measured using the NanoDrop Spectrophotometer.


KEVIN

1. Glycerol Stock

Glycerol stocks of R0040, B0015 J13002, and Pqrr4 were made in order to preserve them long term. The addition of glycerl is important because otherwise the cellular membrane would shear.

2. Plasmid Isolation

To obtain more isolated R0040, B0015, J1302, and Pqrr4, miniprep (plasmid isolation) was done on them.


MANDY

Blog Making

Put together a workable blog and other online accounts for our team to update each other on what they've been up to, with an established schedule and everything :) Now I just have to figure out how to get it onto our wiki, or if I have to transfer each post manually from http://igemcalgary.blogspot.com

PRIMA

Marketing & Sponsorship

After constantly following up with companies over the past few weeks, I successfully recruited Alberta Research Council to sponsor the University of Calgary's iGEM team. They have generously donated %250 dollars this year.

We also edited the June newsletter and discussed it with the rest of the marketing team prior to sending it to all the companies we had contacted (including those who have declined our proposal).


VICKI

Sequencing of what I hope will be LuxOD47A BBK

I prepared a sample of the recently-formed-and-colony-PCR'd-and-NotI-restriction-digested LuxOD47A BBk for sequencing. This was done with BBk sequencing primers, in accordance with the sequencing protocol.