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− | <div class="navbar-header"> <a href="https://2018.igem.org/Team:BJRS_China" target="_top" class="navbar-brand"><img src="https://static.igem.org/mediawiki/2018/7/73/T--BJRS_China--logo.png" width=" | + | <div class="navbar-header"><a href="https://2018.igem.org/Team:BJRS_China" target="_top" class="navbar-brand"><img src="https://static.igem.org/mediawiki/2018/7/73/T--BJRS_China--logo.png" width="120px" height="120px" style="position:relative;top:0px;left:90%;" class="img-fluid rounded"><img src="https://static.igem.org/mediawiki/2018/b/b3/T--BJRS_China--logo2.jpeg"width="1200px" height="50px"style="position:relative;top:-114px;left:10px;"></a> |
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− | <li><a href=" | + | <li><a href="https://2018.igem.org/Team:BJRS_China/Design">DESIGN</a></li> |
+ | <li><a href="https://2018.igem.org/Team:BJRS_China/Results">RESULTS</a></li> | ||
<li><a href="https://2018.igem.org/Team:BJRS_China/Improve">IMPROVE</a></li> | <li><a href="https://2018.igem.org/Team:BJRS_China/Improve">IMPROVE</a></li> | ||
+ | <li><a href="https://2018.igem.org/Team:BJRS_China/InterLab">INTERLAB</a></li> | ||
+ | <li><a href="https://2018.igem.org/Team:BJRS_China/Demonstrate">DEMONSTRATE</a></li> | ||
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− | <li class="dropdown"> <a href="#" class="dropdown-toggle" data-toggle="dropdown"> | + | <li class="dropdown"> <a href="#" class="dropdown-toggle" data-toggle="dropdown"> ACHIEVEMENTS <b class="caret"></b> </a> |
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− | <li><a href="https://2018.igem.org/Team:BJRS_China/ | + | <li><a href="https://2018.igem.org/Team:BJRS_China/Parts">PARTS</a></li> |
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− | <li><a href=" | + | <li><a href="https://2018.igem.org/Team:BJRS_China/Notebooks">NOTEBOOKS</a></li> |
+ | <li><a href="https://2018.igem.org/Team:BJRS_China/Protocol">PROTOCOL</a></li> | ||
+ | <li><a href="https://2018.igem.org/Team:BJRS_China/Acknowledgement">ACKNOWLEDGEMENT</a></li> | ||
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<h1>Preparation</h1> | <h1>Preparation</h1> | ||
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<p> | <p> | ||
<font size="5"> | <font size="5"> | ||
− | + | We participated in answering the following question during the InterLab study of this year: | |
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<font color = "orange" size = "4" face = "serif"> | <font color = "orange" size = "4" face = "serif"> | ||
− | + | Can we reduce lab-to-lab variability in fluorescence measurements by normalizing to absolute cell count or colony-forming units (CFUs) instead of OD? | |
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<h2>Materials</h2> | <h2>Materials</h2> | ||
− | <p>DNA/Plasmids</p> | + | <p><font size="4">DNA/Plasmids</p> |
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<li>Negative Control: BBa_R0040</li> | <li>Negative Control: BBa_R0040</li> | ||
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<li>Test Device 6: BBa_J364009</li> | <li>Test Device 6: BBa_J364009</li> | ||
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− | <p>Apparatus</p > | + | <p><font size="4">Apparatus</font></p > |
<ul> | <ul> | ||
<li>96 well plates (provided by Peking University)</li> | <li>96 well plates (provided by Peking University)</li> | ||
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<li>Pipettes</li> | <li>Pipettes</li> | ||
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<li>LUDOX CL-X</li> | <li>LUDOX CL-X</li> | ||
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<h2>Protocols</h2> | <h2>Protocols</h2> | ||
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<h2>I.Calibrations</h2> | <h2>I.Calibrations</h2> | ||
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− | <p>OD<sub>600</sub> reference point:</p> | + | <p><font size = "3"><b>OD<sub>600</sub> reference point:</b></font></p> |
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− | + | <img src="https://static.igem.org/mediawiki/2018/a/af/T--BJRS_China--OD600_Reference_Point.png" alt = "OD<sub>600</sub> Reference Point" style = "width: 50%"> | |
+ | <figcaption><font size = "1"> | ||
+ | Table 1. OD600 Reference Point.</font> | ||
+ | </figcaption> | ||
+ | </figure> | ||
+ | <p></p> | ||
</div> | </div> | ||
− | <div | + | <div> |
− | <p>Particle Standard Curve</p> | + | <p><font size = "3"><b>Particle Standard Curve</b></font></p> |
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− | + | <img src = "https://static.igem.org/mediawiki/2018/e/ef/T--BJRS_China--Particle_Standard_Curve.jpg" alt = "Particle Standard Curve" style = "width: 70%"> | |
+ | <figcaption><font size = "1"> | ||
+ | Figure 1. Particle Standard Curve.</font> | ||
+ | </figcaption> | ||
+ | </figure> | ||
</div> | </div> | ||
− | <div | + | <div> |
− | <p>Particle Standard Curve(log scale)</p> | + | <p><font size = "3"><b>Particle Standard Curve(log scale)</b></font></p> |
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− | + | <img src = "https://static.igem.org/mediawiki/2018/d/dd/T--BJRS_China--Particle_Standard_Curve%28log_scale%29.jpg" alt = "Particle Standard Curve(log scale)" style = "width: 70%"> | |
+ | <figcaption><font size = "1"> | ||
+ | Figure 1. Particle Standard Curve(log scale).</font> | ||
+ | </figcaption> | ||
+ | </figure> | ||
</div> | </div> | ||
− | <div | + | <div> |
− | <p>Fluorescein Standard Curve</p> | + | <p><font size = "3"><b>Fluorescein Standard Curve</b></font></p> |
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+ | <figcaption><font size = "1"> | ||
+ | Figure 2. Fluorescein Standard Curve.</font> | ||
+ | </figcaption> | ||
+ | </figure> | ||
</div> | </div> | ||
− | <div | + | <div> |
− | <p>Fluorescein Standard Curve(log scale)</p> | + | <p><font size = "3"><b>Fluorescein Standard Curve(log scale)</b></font></p> |
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+ | <figcaption><font size = "1"> | ||
+ | Figure 2. Fluorescein Standard Curve(log scale).</font> | ||
+ | </figcaption> | ||
+ | </figure> | ||
</div> | </div> | ||
<h2>II.Cell Measurements</h2> | <h2>II.Cell Measurements</h2> | ||
− | <div | + | <div> |
− | <p><b>Fluorescence Raw</b></p> | + | <p><font size = "3"><b>Fluorescence Raw</b></font></p> |
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− | + | <img src = "https://static.igem.org/mediawiki/2018/0/06/T--BJRS_China--Fluorescence_Raw_0_hour.jpg" alt = "Fluorescence at 0h" style = "width: 80%"> | |
+ | <figcaption><font size = "1"> | ||
+ | Table 2. Raw Plate Reader Measurements of Fluorescence Raw at 0 hour.</font> | ||
+ | </figcaption> | ||
+ | </figure> | ||
</div> | </div> | ||
− | <div class = " | + | <div> |
− | + | <figure class="text-center"> | |
− | + | <img src = "https://static.igem.org/mediawiki/2018/1/1d/T--BJRS_China--Fluorescence_Raw_6_hour.jpg" alt = "Fluorescence at 6h" style = "width: 80%"> | |
+ | <figcaption><font size = "1"> | ||
+ | Table 3. Raw Plate Reader Measurements of Fluorescence Raw at 6 hours.</font> | ||
+ | </figcaption> | ||
+ | </figure> | ||
</div> | </div> | ||
− | <div | + | <div> |
− | <p><b>Abs<sub>600</sub> Raw</b></p> | + | <p><font size = "3"><b>Abs<sub>600</sub> Raw</b></font></p> |
− | + | <figure class="text-center"> | |
− | + | <img src = "https://static.igem.org/mediawiki/2018/5/59/T--BJRS_China--Abs600_Raw_0_hour.jpg" alt = "Abs600 at 0h" style = "width: 80%"> | |
+ | <figcaption><font size = "1"> | ||
+ | Table 4. Raw Plate Reader Measurements of Abs600 Raw at 0 hour.</font> | ||
+ | </figcaption> | ||
+ | </figure> | ||
</div> | </div> | ||
− | <div class = " | + | <div> |
− | + | <figure class="text-center"> | |
− | + | <img src = "https://static.igem.org/mediawiki/2018/8/86/T--BJRS_China--Abs600_Raw_6_hour.jpg" alt = "Abs600 at 6h" style = "width: 80%"> | |
− | + | <figcaption><font size = "1"> | |
− | + | Table 5. Raw Plate Reader Measurements of Abs600 Raw at 6 hours.</font> | |
− | + | </figcaption> | |
− | + | </figure> | |
− | + | </div> | |
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Latest revision as of 23:54, 17 October 2018
InterLab
Preparation
We participated in answering the following question during the InterLab study of this year:
Can we reduce lab-to-lab variability in fluorescence measurements by normalizing to absolute cell count or colony-forming units (CFUs) instead of OD?
Materials
DNA/Plasmids
- Negative Control: BBa_R0040
- Positive Control: BBa_I20270
- Test Device 1: BBa_J364000
- Test Device 2: BBa_J364001
- Test Device 3: BBa_J364002
- Test Device 4: BBa_J364007
- Test Device 5: BBa_J364008
- Test Device 6: BBa_J364009
Apparatus
- 96 well plates (provided by Peking University)
- Plate reader
- Foil covered 50 ml tube
- Eppendorf tubes
- Pipettes
Materials
- LUDOX CL-X
- Silica beads
- Fluorescein
- Phosphate buffered saline
- LB media
- Chloramphenicol
- LB plates
- distilled water
Protocols
We followed the protocol provided by iGEM HQ so that inter-laboratory errors can be reduced. Protocols we used can be found here:
2018 InterLab Plate Reader ProtocolHelp: Protocols/Transformation
Results
I.Calibrations
OD600 reference point:
![OD<sub>600</sub> Reference Point](https://static.igem.org/mediawiki/2018/a/af/T--BJRS_China--OD600_Reference_Point.png)
Particle Standard Curve
![Particle Standard Curve](https://static.igem.org/mediawiki/2018/e/ef/T--BJRS_China--Particle_Standard_Curve.jpg)
Particle Standard Curve(log scale)
![Particle Standard Curve(log scale)](https://static.igem.org/mediawiki/2018/d/dd/T--BJRS_China--Particle_Standard_Curve%28log_scale%29.jpg)
Fluorescein Standard Curve
![Fluorescein Standard Curve](https://static.igem.org/mediawiki/2018/9/95/T--BJRS_China--Fluorescein_Standard_Curve.png)
Fluorescein Standard Curve(log scale)
![Fluorescein Standard Curve(log scale)](https://static.igem.org/mediawiki/2018/9/97/T--BJRS_China--Fluorescein_Standard_Curve%28log_scale%29.png)
II.Cell Measurements
Fluorescence Raw
![Fluorescence at 0h](https://static.igem.org/mediawiki/2018/0/06/T--BJRS_China--Fluorescence_Raw_0_hour.jpg)
![Fluorescence at 6h](https://static.igem.org/mediawiki/2018/1/1d/T--BJRS_China--Fluorescence_Raw_6_hour.jpg)
Abs600 Raw
![Abs600 at 0h](https://static.igem.org/mediawiki/2018/5/59/T--BJRS_China--Abs600_Raw_0_hour.jpg)
![Abs600 at 6h](https://static.igem.org/mediawiki/2018/8/86/T--BJRS_China--Abs600_Raw_6_hour.jpg)