iGEM 2014 — Team The_Tech_Museum

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# Team:The Tech Museum/Notebook

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| Notebook | | |
| **Timeline of major events:**  * May: Basic concept research and design * June: Software development initiated; plasmid design started and promoter-RBS’s picked * July: Wetlab testing of possible color protein combinations * August 10: Design of complete tri-color plasmids finalized * August 24: Assembly of tri-color plasmid pools completed (DNA2.0) * September 10: Tri-color plasmids optimized on museum wetlab setup * September 29: Software fully integrated with hardware on mobile exhibit * September 30 - October 3: Prototyping of mobile exhibit with museum staff and visitors * October 6 - October 17: Data collection on the museum floor with visitor!   **Biology Details:**  Design of tri-color plasmid pool    * 9 promoter-rbs pairs of varying strength from Kosuri et al paper (2013):  |  |  |  | | --- | --- | --- | |  | Promoter - RBS Pairs | Sequence |  | 1 | BBa\_J23117 - BBa\_J61112 | TTGACAGCTAGCTCAGTCCTAGGGATTGTGCTAGCCAATCTCTAGAGAAAGAGGTGACATAC | | 2 | BBa\_J23104 - BBa\_J61107 | TTGACAGCTAGCTCAGTCCTAGGTATTGTGCTAGCTTACGTCTAGAGAAAGAAGAGACTCAC | | 3 | apFAB78 - apFAB954 | TTGACATTTATCCCTTGCGGCGATAGATTTAACGTATGACGGATCTTAATCTAGCTCAGGACAATTT | | 4 | apFAB76 - apFAB927 | TTGACATTTATCCCTTGCGGCGATATAATAGATATCTTAATCTAGCCCGGGAGTTTTTTCATTCCGGATCTTAATCTAGCTGGGGACTGTTT | | 5 | apFAB70 - apFAB844 | TTGACATCGCATCTTTTTGTACCTATAATGTGTGGATAGAGTATCTTAATCTAGCAGGGGACACTTT | | 6 | BBa\_J23104 - apFAB909 | TTGACAGCTAGCTCAGTCCTAGGTATTGTGCTAGCTTACGATCTTAATCTAGCGAAGGATAGTTT | | 7 | apFAB45 - apFAB901 | AAAAAGAGTATTGACTTCGCATCTTTTTGTACCTATAATGTGTGGATAGCGG | | 8 | apFAB92 - apFAB863 | AAAAAATTTATTTGCTTTCAGGAAAATTTTTCTGCATAATTATTTCATGGAGCATCTTAATCTAGCGGGGGAGCGTTT | | 9 | apFAB71 - salis-4-10 | TTGACATCGCATCTTTTTGTACCTATAATAGATTCATGATGAAATCTCTTTTATCAAATATAAGCAGGAT |    Selection of colored reporter proteins    * Co-transformation testing of multiple color combinations of proteins from DNA2.0 * Transformations on Kanamycin and IPTG ![](https://static.igem.org/mediawiki/2014/6/6c/Tech_Museum_Chromo-Testing.png)   ![](https://static.igem.org/mediawiki/2014/1/12/Tech_Museum_Fluoro-Testing.png)   Final tri-color plasmid designs, assembled by DNA2.0:  Chromogenic plasmid pool    * Blitzen (blue)  * Kringle (yellow) * Paprika (red)  Fluorescent plasmid pool   * CindyLouCFP (400/495) * KringleYFP (520/542) * PaprikaRFP (554/590)  Optimization of plasmid pools with visitor-accessible museum wetlab set up. Transformation condition:   * 6cm plates * 400 ug/ml Amp * 0.3 ul unamplified plasmid pool in 100ul CaCl2 * 20 ul competent bacteria * Current visitor wetlab transformation protocol: 30 sec on ice, 40 sec heat shot at 42 degrees C * Chromogenic pool maturation time ~ 5 days 37 degrees C * Fluoroescent pool maturation time ~ 3 days 37 degrees C * Low copy fluorescent plasmid pool gave more reliable results with most color variety   **Safety:**  We did not use any dangerous organisms or reagents. Transformation of bacteria was done in the musuem’s existing wetlab setup. | | |  |  |

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