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Revision as of 15:52, 4 October 2015

iGEM logo
Introduction
Antibody issue
PAST
Process
Results
Practices
What is Next

"I've never managed to keep a journal longer than two weeks." - J. K. Rowling

Complete Journal

All our successes and major events is highlighted in the following. We also listed the most important iGEM dates as well as protocols and primer designs. Feel free to click through the boxes, check out our primer designs or read through the protocols for the Lab. The protocols can be accessed by clicking on the protocol number.

Major events
Month: Date: Event:
Febuary 23.02.15 First meeting
March 20.03.15-
22.03.15		 Trip to Langeland
24.03.15 Project idea chosen
25.03.15 Visiting Knud Villy Christiansen on TEK
April 01.04.15 First day in Lab
10.04.15 CFO Easter Party
15.04.15 Lasagna night
24.04.15-
26.04.15		 Danish Meetup
July 24.07.15-
26.07.15		 Nordic Jamboree
28.07.15 Presentation for UNF-camp
August 03.08.15 Visiting animal testing facility
05.08.15 Wiki and Wine
September 14.09.15 iGEM Alumne - Dinner with the old iGEM members
16.09.15 Sleepover 1.0 in the "Lab"
19.09.15 Wiki-freeze Brunch
20.09.15 Movie Night: Back to the Future!
Lab successes

Month:

Date:

Succes:
April 02.04.15
  • Created our first bio-bricks T18 and T25: MG1655:pSB1C3-T18, MG1655:pSB1K3-T18, MG1655:pSB1C3-T25 and MG1655:pSB1K3-T25
    		•
    May 11.05.15
  • Test with GFP using FAX 11/05-2015: MG1655:pSB1C3-T18-Linker-GFP and MG1655:pSB1C3-T25-Linker-GFP
    		•
    August 17.08.15
  • Knockout creation: MG1655:ΔCyA
    		•
    19.08.15
  • Final construct of T25: MG1655:pSB1K3-T25-Target
    		•
    27.08.15
  • Final construct of T18: MG1655:pSB1A3-T18-Linker-Scaffold-Library-Scaffold-3xFLAG
    		•
    28.08.15
  • Interlab study, completed with FAX on the constructs: TOP10:pSB1C3-J23101-I13504, TOP10:pSB1C3-J23106-I13504, TOP10:pSB1C3-J23117-I13504
    		•
    September 04.09.15
  • Receiving a peptide aptamer library of our own design from IDT
    		•
    08.09.15
  • Positive control: MG1655: pSB1C3-T18-Linker-Leucin Zipper + pSB1K3-T25-Linker-Leuzine Zipper
    		•
    17.09.15
  • Caraktezaition: Northern blot on MG1655:pSB1C3-PcstA -GFP
    		•
    18.09.15
  • First peptide aptamer screening??
    		•
    iGEM deadlines

    Month:

    Date:

    Deadline:
    March 31.03.15 Team registration closes
    July 15.07.15 Project Description
    31.07.15 Giant Jamboree registration closes
    August 07.08.15 Track Selection
    07.08.15 Project title and abstract
    28.08.15 Interlab Study data due
    September 18.09.15 Parts Submission
    18.09.15 Wiki-freeze
    24.09.15-
    28.09.15    		 Giant Jamboree
    Protocols

    Number:

    Name:

    Description
    000 Template
  • Template for all the protocols made by SDU iGEM 2015.
    		•
    001 Linker GFP
  • Assembly of a biobrick that contains the Linker-GFP gene.
    		•
    002 Control of Leucine Zipper
  • To conjugate a leucin zipper to the T18 and T25 domain as used in the bacterial
    two-hybrid system and to furthermore verify that the bacterial two-hybrid system
    works
    		•
    003 Interlab Study
  • Measurement of GFP under influence of different promoters.
    		•
    004 cAMP Indicator
  • To create a brick that can indicate the activation of CyaA in the two hybrid
    system, by colour expression when increased presence of cAMP.
    		•
    005 CyaA Knockout
  • To create a knockout of the CyaA gene in MG1655. This is done to get a bacterial
    strain which we can use to use and test the Two-Hybrid System.
    		•
    006 Characterization of PcstA
  • To better characterize the cAMP­inducable promotor PcstA. Three experiments are designed and executed in this protocol.
    		•
    007 The Two Hybrid System
  • To construct two bricks that interact with each other allowing CyaA activation. The activation only occurs if the proteins bound to CyaA interact with each other. This interaction allows CyaA to produce cAMP from ATP.
    		•
    008 Peptide Aptamer Screening
  • To screen for peptide aptamers using the two­hybrid system
    		•
    Primers

    Number:

    Name:

    Sequence:
    001 pKT25_BB_F 5’-ATATGAATTCGCGGCCGCTTCTAGAGCTGGCACGACAGGTTTCCCG-3’
    002 pKT25_BB_R 5’-ATATCTGCAGCGGCCGCTACTACTAGGATCCAGCCCGCCGCGT-3’
    003 pUT18_BB_R 5’-ATATCTGCAGCGGCCGCTACTAGTAGGATCCGCGTTCCACTGCGCCC-3’
    004 Linker-GFP_F 5’-CGCTTCTAGCGGGATCCGAAAATTTGTATTTTCAATCTGGTATGCGTAAAGGAGAAGAACTTTTCACT
    GG-3’
    005 Con-DsRed_F 5’-CGCTTCTAGAGTTTACGGCTAGCTCAGTCCTAGGTATAGTGCTAGCAAAGAGGAGAAATACTAGATGG
    ATAGCACTGAGAACGTCATCAAGC-3’
    006 Con-DsRed_R 5’-ATATCTGCAGCGGCCGCTACTAGTAGCGAAAAAACCCCGCCGAAGCGGGGTTTTTTGCGTTATTACTG
    GAACAGGTGGTGGCGG-3’
    007 GFP-mCherry_R/
    GFP-DsRed_R    		 5’-GAGCTAGCCGTAAACTAGTATATAAACGCAGAAAGGCCCACCC-3’
    008 GFP-mCherry_F/
    GFP-DsRed_F    		 5’-CTGCGTTTATATACTAGTTTACGGCTAGCTCAGTCCTAGGG-3’
    009 Con-mCherry_R 5’-ATATCTGCAGCGGCCGCTACTAGTATATAAACGCAGAAAGGCCCACCC-3’
    010 Con-mCherry_F 5’-CGCTTCTAGCGTTTACGGCTAGCTCAGTCCTAGGTATAGTGCTAGCAAAGAGGAGAAATACTAGATGG
    TGAGCAAGG-3’
    011 cyaA_F 5’-GAGGTCTGGCAGTGGATCCT-3’
    012 cyaA_R 5’-GAGTGGATTTGTGATCGCAGCG-3’
    013 CyaA_del_F 5’-GGAATTTACAGAGAATAAACGGTGCTACACTTGTATGTAGCGCATCTTTCTGTGTAGGCTGGAGCTGC
    TTC-3’
    014 cyaA_del_R 5’-GTTTCCGCTAAGATTGCATGCCGGATAAGCCTCGCTTTCCGGCACGTTCACATATGAATATCCTCCTT
    AG-3’
    015 FWLeuZ-F 5’-CCCCGGATCCAAACCGTTCCAGTGCCG-3’
    016 RSLeuZ-R 5’-ATATCTGCAGCGGCCGCTACTAGTAGAGATCTACACTAGCACTATCAGAGTTATTAAGC-3’
    017 iGEM VF2 5’-CCACCTGACGTCTAAGAAAC-3’
    018 iGEM VR 5’-ATTACCGCCTTTGAGTGAGC-3’
    019 Zip_F 5’-CGCTTCTAGAGGGATCCGAAAATTTGTATTTTCAATCTGGTATGAAACAGCTGGAAGACAAAGTTGA-3’
    020 Zip_R 5’-ATATCTGCAGCGGCCGCTACTAGTAACGTTCACCAACCAGTTTTTTCAGA-3’
    021 H1F_CyaA_del 5’-GCATCCAGACTGTCATAGTTATTGGAC-3’
    022 H1R_CyaA_del 5’-GAAAGATGCGCTACATACAAGTGTAGC-3’
    023 H2F_CyaA_del 5’-TGAACGTGCCGGAAAGCGAG-3’
    024 H2R_CyaA_del 5’-AAGTGAGGAAATGCTGCGAGTATAAC-3’
    025 GFP_LVA_F 5’-GCGCCGTCTAGAAAAGAGGAGAAATACTAGATGCGTAAAGGAGAAGAACTTTTCACT-3’
    026 GFP_LVA_F 5’-GCCGGACTGCAGCGGCCGCTACTAGTATTAAGCTACTAAAGCGTAGTTTTCGTCG-3’

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