Difference between revisions of "Team:UST Beijing/InterLab"

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      <h1 class="title">Interlab</h1>
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      <h1 class="intro">
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      <span  class="hue">InterLab Study</span>was very helpful before the formal experiment started. By following InterLab protocols, we can easily establish standardized, repeatable and reliable measurement tools. The aim of this year’s InterLab study was to establish standard curves of particles and fluorescein using a plate reader and measure GFP fluorescence. </br>
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      Follow the protocol given by iGEM,we <span>completed</span> the required experiments.What's more,we helped high school iGEMers with experimental equipments and instructions.<br/>
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      <h1>Here are details——<span>Materials/Discussion/Conclusion</span>. Results are as follows.</h1>
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    <h3><span>Materials</span></h3>
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<h4>- Plate reader:  PerkinElmer EnSpire</br>
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- 96 well plates (transparent plates with clear flat bottom)</br>
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    - Devices: Positive control:BBa_I20270</br>
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Negative control: BBa_R0040</br>
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        Device 1: BBa_J364000
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        Device 2: BBa_J364001.
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        Device 3: BBa_J364002
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        Device 4: BBa_J364007
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        Device 5: BBa_J364008
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    Device 6: BBa_J364009</br>
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- Fluorescein (provided in kit)</br>
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    - 10ml 1xPBS pH 7.4-7.6</br> 
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    (phosphate buffered saline)</br>
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- 300 μL Silica beads </br>
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    - Microsphere suspension
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    </br>(provided in kit, 4.7x10^8 microspheres)
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        - 1ml LUDOX CL-X (provided in kit)</br>
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        - ddH2O </h4>
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    <h3><span>Conclusion</span></h3>
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            <h4>Both Device 2 and Device 4 showed higher fluorescence values than other Devices, and Device 2 is the best. The fluorescence result of Device 3 is the lowest, still higher than negative control though.</h4>
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    <h3><span>Discussion</span></h3>
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        <h4>Figure 1 demonstrated that under 5uM Fluorescein, the standard curve showed a similar polynomial shape, but the whole curve didn’t. Therefore, we speculated that 10uM Fluorescein might beyond the range of the equipment, which meant the concentration of fluorescein was too high to acquire reliable data. According to Figure 2, the log graph of fluorescein standard curve showed a highly similar linear shape.</br>Particle standard curve (Figure 3 and Figure 4) demonstrated the same problem above (beyond the scale). According to figure 4, particle count under 5×10^7 showed a highly similar linear shape.</br>Table 1 demonstrated values of Net Fluorescein a.u. From data of 6 hours. We could notice that after 6-hour growth, Device 2 (BBa_J364001) showed a series of high values which were even higher than Positive control. Furthermore, according to Table 2, the Net Abs600 values of them just had little difference, which meant within the range of the errors allowed, the growth of Positive control and Device 2 are similar. Additionally, Device 4 also showed higher value, however, the 0-hour Net fluorescein a.u. values of Device 4 were higher than any other Devices and replicates of colony 1 of Device4 were not very good. Thus, we only compared colony 2 of Device 2 and Device 4, although the initial fluorescein values of device 4 are much higher, they became lower after 6 hours.</h4>
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                              <a href="https://static.igem.org/mediawiki/2018/4/45/T--UST_Beijing--interlabpic1.jpg"><span>Interlab</span></a><br/>
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                              <a href="https://static.igem.org/mediawiki/2018/1/15/T--UST_Beijing--interlabpic2.jpg"><span>with highschool igemers</span></a><br/>
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<h1>InterLab</h1>
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<h3>Bronze Medal Criterion #4</h3>
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<p><b>Standard Tracks:</b> Participate in the Interlab Measurement Study and/or obtain new, high quality experimental characterization data for an existing BioBrick Part or Device and enter this information on that part's Main Page in the Registry. The part that you are characterizing must NOT be from a 2018 part number range.
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For teams participating in the <a href="https://2018.igem.org/Measurement/InterLab">InterLab study</a>, all work must be shown on this page.
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Revision as of 22:10, 8 October 2018

<!DOCTYPE html> flati

Materials

- Plate reader: PerkinElmer EnSpire
- 96 well plates (transparent plates with clear flat bottom)
- Devices: Positive control:BBa_I20270
Negative control: BBa_R0040
Device 1: BBa_J364000 Device 2: BBa_J364001. Device 3: BBa_J364002 Device 4: BBa_J364007 Device 5: BBa_J364008 Device 6: BBa_J364009
- Fluorescein (provided in kit)
- 10ml 1xPBS pH 7.4-7.6
(phosphate buffered saline)
- 300 μL Silica beads
- Microsphere suspension
(provided in kit, 4.7x10^8 microspheres) - 1ml LUDOX CL-X (provided in kit)
- ddH2O

Conclusion

Both Device 2 and Device 4 showed higher fluorescence values than other Devices, and Device 2 is the best. The fluorescence result of Device 3 is the lowest, still higher than negative control though.

Discussion

Figure 1 demonstrated that under 5uM Fluorescein, the standard curve showed a similar polynomial shape, but the whole curve didn’t. Therefore, we speculated that 10uM Fluorescein might beyond the range of the equipment, which meant the concentration of fluorescein was too high to acquire reliable data. According to Figure 2, the log graph of fluorescein standard curve showed a highly similar linear shape.
Particle standard curve (Figure 3 and Figure 4) demonstrated the same problem above (beyond the scale). According to figure 4, particle count under 5×10^7 showed a highly similar linear shape.
Table 1 demonstrated values of Net Fluorescein a.u. From data of 6 hours. We could notice that after 6-hour growth, Device 2 (BBa_J364001) showed a series of high values which were even higher than Positive control. Furthermore, according to Table 2, the Net Abs600 values of them just had little difference, which meant within the range of the errors allowed, the growth of Positive control and Device 2 are similar. Additionally, Device 4 also showed higher value, however, the 0-hour Net fluorescein a.u. values of Device 4 were higher than any other Devices and replicates of colony 1 of Device4 were not very good. Thus, we only compared colony 2 of Device 2 and Device 4, although the initial fluorescein values of device 4 are much higher, they became lower after 6 hours.

Interlab
with highschool igemers
UST_Beijing © .igem.org