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+ | <meta charset="utf-8"> | ||
+ | <title>BJRS</title> | ||
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+ | <nav class="navbar navbar-default" role="navigation"> | ||
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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="100px" height="100px" style="position:relative;top:-12px;left:20px;" class="img-fluid rounded"><img src="https://static.igem.org/mediawiki/2018/8/8b/HQ_page_logo.jpg" width="100px" height="100px" style="position:relative;top:-115px;left: -125px" class="img-fluid rounded"></a> | ||
</div> | </div> | ||
+ | <div> | ||
+ | <ul class="nav navbar-nav"> | ||
+ | <li class="dropdown"> <a href="#" class="dropdown-toggle" data-toggle="dropdown"> PROJECT <b class="caret"></b> </a> | ||
+ | <ul class="dropdown-menu"> | ||
+ | <li><a href="https://2018.igem.org/Team:BJRS_China">BACKGROUND</a></li> | ||
+ | <li><a href="#">PROJECT</a></li> | ||
+ | <li><a href="https://2018.igem.org/Team:BJRS_China/Improve">IMPROVE</a></li> | ||
+ | </ul> | ||
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+ | <li><a href="#">EXPERIMENT</a></li> | ||
+ | <li><a href="https://2018.igem.org/Team:BJRS_China/InterLab">INTERLAB</a></li> | ||
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+ | </ul> | ||
+ | </li> | ||
+ | <li><a href="#">SAFETY</a></li> | ||
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+ | <li><a href="https://2018.igem.org/Team:BJRS_China/Collaborations">COLLABORATION</a></li> | ||
+ | </ul> | ||
+ | </li> | ||
+ | <li class="dropdown"> <a href="#" class="dropdown-toggle" data-toggle="dropdown"> TEAM <b class="caret"></b> </a> | ||
+ | <ul class="dropdown-menu"> | ||
+ | <li><a href="#">MEMBERS</a></li> | ||
+ | <li><a href="#">SCHOOLS</a></li> | ||
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+ | </div> | ||
+ | </nav> | ||
+ | <main> | ||
+ | <section class = "interlab"> | ||
+ | <!--Title--> | ||
+ | <h1 style = "text-align: center">InterLab</h1> | ||
− | < | + | <hr class="line"> |
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
+ | <div> | ||
+ | <h1>Preparation</h1> | ||
+ | <h2>What Is Interlab?</h2> | ||
+ | <p> <br> | ||
+ | <br> | ||
+ | This is the 5th year of InterLab and we have the following question: | ||
+ | <br> | ||
+ | <b> | ||
+ | <font color = "orange" size = "5" 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? | ||
+ | </font> | ||
+ | </b> | ||
+ | </p> | ||
+ | |||
+ | </div> | ||
+ | |||
+ | <div> | ||
+ | <h2>Materials</h2> | ||
+ | <p>DNA/Plasmids</p> | ||
+ | <ol> | ||
+ | <li>Negative Control: BBa_R0040</li> | ||
+ | <li>Positive Control: BBa_I20270</li> | ||
+ | <li>Test Device 1: BBa_J364000</li> | ||
+ | <li>Test Device 2: BBa_J364001</li> | ||
+ | <li>Test Device 3: BBa_J364002</li> | ||
+ | <li>Test Device 4: BBa_J364007</li> | ||
+ | <li>Test Device 5: BBa_J364008</li> | ||
+ | <li>Test Device 6: BBa_J364009</li> | ||
+ | </ol> | ||
+ | <p>Apparatus</p > | ||
+ | <ul> | ||
+ | <li>96 well plates (provided by Peking University)</li> | ||
+ | <li>Plate reader</li> | ||
+ | <li>Foil covered 50 ml tube</li> | ||
+ | <li>Eppendorf tubes</li> | ||
+ | <li>Pipettes</li> | ||
+ | </ul> | ||
+ | <p>Materials</p> | ||
+ | <ul> | ||
+ | <li>LUDOX CL-X</li> | ||
+ | <li>Silica beads</li> | ||
+ | <li>Fluorescein</li> | ||
+ | <li>Phosphate buffered saline</li> | ||
+ | <li>LB media</li> | ||
+ | <li>Chloramphenicol</li> | ||
+ | <li>LB plates</li> | ||
+ | <li>distilled water</li> | ||
+ | </ul> | ||
+ | </div> | ||
+ | |||
+ | <div> | ||
+ | <h2>Protocols</h2> | ||
+ | <div> | ||
+ | <p> We followed the protocol provided by iGEM HQ so that inter-laboratory errors can be reduced. Protocols we used can be found here:</p> | ||
+ | <a href = "https://static.igem.org/mediawiki/2018/0/09/2018_InterLab_Plate_Reader_Protocol.pdf"> 2018 InterLab Plate Reader Protocol</a><br> | ||
+ | <a href = "http://parts.igem.org/Help:Protocols/Transformation">Help: Protocols/Transformation</a> | ||
+ | </div> | ||
+ | <div> | ||
+ | <h1>Experiment Process</h1> | ||
+ | <p></p> | ||
+ | <div> | ||
+ | <p></p> | ||
+ | <img src = "" alt = "Open Distribution Kit" style = "width: 30%"> | ||
+ | </div> | ||
+ | <div> | ||
+ | <p></p> | ||
+ | <img src = "" alt = "Add silica beads to 96 well plate" style = "width: 30%"> | ||
+ | </div> | ||
+ | <div> | ||
+ | <p></p> | ||
+ | |||
+ | </div> | ||
+ | <div> | ||
+ | <p></p> | ||
+ | </div> | ||
+ | </div> | ||
+ | |||
+ | <div> | ||
+ | <h1>Results</h1> | ||
+ | <p><i>The results are shown in the tables and figures.</i></p> | ||
+ | <h2>I.Calibrations</h2> | ||
+ | <div class="inlabdiv"> | ||
+ | <p>OD<sub>600</sub> reference point:</p> | ||
+ | <img src="https://static.igem.org/mediawiki/2018/a/af/T--BJRS_China--OD600_Reference_Point.png" alt = "OD600 Reference Point" style = "width: 30%"><br> | ||
+ | <font size = "1">Table 1. OD600 Reference Point.</font> | ||
+ | </div> | ||
+ | <div class="inlabdiv"> | ||
+ | <p>Particle Standard Curve</p> | ||
+ | <img src = "https://static.igem.org/mediawiki/2018/e/ef/T--BJRS_China--Particle_Standard_Curve.jpg" alt = "Particle Standard Curve" style = "width: 50%"><br> | ||
+ | <font size = "1">Figure 1. Particle Standard Curve.</font> | ||
+ | </div> | ||
+ | <div class="inlabdiv"> | ||
+ | <p>Particle Standard Curve(log scale)</p> | ||
+ | <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: 50%"><br> | ||
+ | <font size = "1">Figure 1. Particle Standard Curve(log scale).</font> | ||
+ | </div> | ||
+ | |||
+ | <div class="inlabdiv"> | ||
+ | <p>Fluorescein Standard Curve</p> | ||
+ | <img src = "" alt = "Fluorescein Standard Curve" style = "width: 50%"><br> | ||
+ | <font size = "1">Figure 2. Fluorescein Standard Curve.</font> | ||
+ | </div> | ||
+ | <div class="inlabdiv"> | ||
+ | <p>Fluorescein Standard Curve(log scale)</p> | ||
+ | <img src = "" alt = "Fluorescein Standard Curve(log scale)" style = "width: 50%"><br> | ||
+ | <font size = "1">Figure 2. Fluorescein Standard Curve(log scale).</font> | ||
+ | </div> | ||
+ | <h2>II.Cell Measurements</h2> | ||
+ | <div class="inlabdiv"> | ||
+ | <p><b>Fluorescence Raw</b></p> | ||
+ | <img src = "https://static.igem.org/mediawiki/2018/0/06/T--BJRS_China--Fluorescence_Raw_0_hour.jpg" alt = "Fluorescence at 0h" style = "width: 70%"><br> | ||
+ | <font size = "1">Table 2. Raw Plate Reader Measurements of Fluorescence Raw at 0 hour.</font> | ||
+ | </div> | ||
+ | <div class = "inlabdiv"> | ||
+ | <img src = "https://static.igem.org/mediawiki/2018/1/1d/T--BJRS_China--Fluorescence_Raw_6_hour.jpg" alt = "Fluorescence at 6h" style = "width: 70%"><br> | ||
+ | <font size = "1">Table 3. Raw Plate Reader Measurements of Fluorescence Raw at 6 hours.</font> | ||
+ | </div> | ||
+ | <div class = "inlabdiv"> | ||
+ | <p><b>Abs<sub>600</sub> Raw</b></p> | ||
+ | <img src = "https://static.igem.org/mediawiki/2018/5/59/T--BJRS_China--Abs600_Raw_0_hour.jpg" alt = "Abs600 at 0h" style = "width: 70%"><br> | ||
+ | <font size = "1">Table 4. Raw Plate Reader Measurements of Abs600 Raw at 0 hour.</font> | ||
+ | </div> | ||
+ | <div class = "inlabdiv"> | ||
+ | <img src = "https://static.igem.org/mediawiki/2018/8/86/T--BJRS_China--Abs600_Raw_6_hour.jpg" alt = "Abs600 at 6h" style = "width: 70%"><br> | ||
+ | <font size = "1">Table 5. Raw Plate Reader Measurements of Abs600 Raw at 6 hours.</font> | ||
+ | </div> | ||
+ | |||
+ | <div> | ||
+ | <h1>Discussion</h1> | ||
+ | <p></p> | ||
+ | <p></p> | ||
+ | </div> | ||
+ | |||
+ | <div> | ||
+ | <h1>Our Thoughts and Feelings</h1> | ||
+ | <p></p> | ||
+ | </div> | ||
+ | </body> | ||
</html> | </html> |
Revision as of 07:12, 5 October 2018
InterLab
Preparation
What Is Interlab?
This is the 5th year of InterLab and we have the following question:
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
Experiment Process
Results
The results are shown in the tables and figures.
I.Calibrations
OD600 reference point:
![OD600 Reference Point](https://static.igem.org/mediawiki/2018/a/af/T--BJRS_China--OD600_Reference_Point.png)
Table 1. OD600 Reference Point.
Particle Standard Curve
![Particle Standard Curve](https://static.igem.org/mediawiki/2018/e/ef/T--BJRS_China--Particle_Standard_Curve.jpg)
Figure 1. Particle Standard Curve.
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)
Figure 1. Particle Standard Curve(log scale).
Fluorescein Standard Curve
Figure 2. Fluorescein Standard Curve.
Fluorescein Standard Curve(log scale)
Figure 2. Fluorescein Standard Curve(log scale).
II.Cell Measurements
Fluorescence Raw
![Fluorescence at 0h](https://static.igem.org/mediawiki/2018/0/06/T--BJRS_China--Fluorescence_Raw_0_hour.jpg)
Table 2. Raw Plate Reader Measurements of Fluorescence Raw at 0 hour.
![Fluorescence at 6h](https://static.igem.org/mediawiki/2018/1/1d/T--BJRS_China--Fluorescence_Raw_6_hour.jpg)
Table 3. Raw Plate Reader Measurements of Fluorescence Raw at 6 hours.
Abs600 Raw
![Abs600 at 0h](https://static.igem.org/mediawiki/2018/5/59/T--BJRS_China--Abs600_Raw_0_hour.jpg)
Table 4. Raw Plate Reader Measurements of Abs600 Raw at 0 hour.
![Abs600 at 6h](https://static.igem.org/mediawiki/2018/8/86/T--BJRS_China--Abs600_Raw_6_hour.jpg)
Table 5. Raw Plate Reader Measurements of Abs600 Raw at 6 hours.