Team:SDU-Denmark/Tour41
"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 | Characterization: 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’ |