Team:ATOMS-Turkiye/Interlab-Study

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Interlab-Study

  • This year, our team was involved in the international scientific experiment study organized by iGEM and declared on the main page of the iGEM website. Considering the promising nature of intensifying international collaboration and its ability to obtain more significant and accurate results, we were delighted to take part in this study and organization.
  • It is important to remind ourselves of the interlab measurement study basics and the expectations from each lab before explaining the kind of process we carried out and what we have accomplished by joining this study.
  • As it is summarized on the iGEM Measurement Study page, participating team must fulfill these requirements:
    • 1. Designing a wiki page about the contribution and the achievements of study.
    2. Constructing the particular BioBricks defined clearly on the page. (The results are shown below.)
    3. Collecting fluorescence data from each of these parts. (The results are sent to iGEM HQ and also shown below.)
    4. Submitting these data through Interlab Study form. (To check out our form: here )
  • Due to achieving all these requirements, we began to work in parallel with the identified tasks:
    • a. Existing device: BBa_I20260 (J23101-B0032-E0040-B0015) in the pSB3K3 vector.
    b. New device to be built by the iGEM team: BBa_J23101 + BBa_E0240 (B0032-E0040-B0015), must be built in the pSB1C3 backbone
    c. New device to be built by the iGEM team: BBa_J23115 + BBa_E0240 (B0032-E0040-B0015), must be built in the pSB1C3 backbone
  • In order to present the results of each part, we separated the experimental approach into four sections:
    • - PRE-EXPERIMENTAL PREPARATION: We planned to use E.coli BL21 (DE3) strain in order to obtain high amounts of protein yield enabling us to calculate the fluorescence emission. Prior for the cloning process, we prepared competent cells using this protocol. LINK
    - FOR CLONING: BBA_I20260 We obtain this part from its iGEM kit plate location and transformed into our prepared BL21 (DE3) competent cells. A detailed gene cloning protocol can be found on this link. LINK
    - FOR CONSTRUCTING: PART BBA_J23101 + BBA_E0240 These parts are collected from iGEM kit plates also and the same cloning protocol LINK is performed afterwards. Successfully cloned parts are ligated according to this protocol. LINK
    - FOR CONSTRUCTING: BBA_J23115 + BBA_E0240
  • These parts are also collected from iGEM kit plates and the same cloning protocol LINK is performed afterwards. Successfully cloned parts are ligated according to this protocol. LINK
    • After the cloning phases, the protein consumption of bacteria cultures is obtained and the protein concentrations are measured as follows:
  • Afterwards, our fluorescence measurement protocol LINK is performed to measure the fluorescence emission. According to the table below, GFP excitation and emission are expected to be observed between these parameters measured by these filters.
  • Our graphic result can be seen below allowing comparisons with the table to be made:
  • The graphs show that the excitation and emission levels of GFP are high enough to say that it is significantly present in the cultured media. Additionally, blank samples have shown no presence of fluorescence in the protein concentration and emission experiments.
    • Design BioBricks

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