Difference between revisions of "Team:BJRS China/InterLab"

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            <li><a href="https://2018.igem.org/Team:BJRS_China">BACKGROUND</a></li>
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            <li><a href="#">PROJECT</a></li>
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<main>
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      <section class = "interlab">
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        <!--Title-->
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      <h1 style = "text-align: center">InterLab</h1>
  
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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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<br><br>
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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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</p>
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</div>
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<div>
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<h1>Preparation</h1>
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                        <h2>What Is Interlab?</h2>
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<p> <br>
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<br>
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This is the 5th year of InterLab and we have the following question:
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<br>
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<b>
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<font color = "orange" size = "5" face = "serif">
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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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</font>
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</b>
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</p>
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</div>
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<div>
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        <h2>Materials</h2>
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<p>DNA/Plasmids</p>
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<ol>
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<li>Negative Control: BBa_R0040</li>
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<li>Positive Control: BBa_I20270</li>
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<li>Test Device 1: BBa_J364000</li>
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<li>Test Device 2: BBa_J364001</li>
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<li>Test Device 3: BBa_J364002</li>
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<li>Test Device 4: BBa_J364007</li>
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<li>Test Device 5: BBa_J364008</li>
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<li>Test Device 6: BBa_J364009</li>
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</ol>
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<p>Apparatus</p >
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<ul>
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<li>96 well plates (provided by Peking University)</li>
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<li>Plate reader</li>
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<li>Foil covered 50 ml tube</li>
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<li>Eppendorf tubes</li>
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<li>Pipettes</li>
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</ul>
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<p>Materials</p>
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<ul>
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<li>LUDOX CL-X</li>
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<li>Silica beads</li>
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<li>Fluorescein</li>
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<li>Phosphate buffered saline</li>
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<li>LB media</li>
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<li>Chloramphenicol</li>
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<li>LB plates</li>
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<li>distilled water</li>
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</ul>
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</div>
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<div>
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<h2>Protocols</h2>
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<div>
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<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>
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<a href = "https://static.igem.org/mediawiki/2018/0/09/2018_InterLab_Plate_Reader_Protocol.pdf"> 2018 InterLab Plate Reader Protocol</a><br>
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<a href = "http://parts.igem.org/Help:Protocols/Transformation">Help: Protocols/Transformation</a>
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</div>
  
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        <div>
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<h1>Experiment Process</h1>
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<p></p>
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<div>
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<p></p>
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<img src = "" alt = "Open Distribution Kit" style = "width: 30%">
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</div>
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<div>
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<p></p>
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<img src = "" alt = "Add silica beads to 96 well plate" style = "width: 30%">
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</div>
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<div>
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<p></p>
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</div>
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<div>
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<p></p>
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</div>
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</div>
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<div>
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<h1>Results</h1>
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<p><i>The results are shown in the tables and figures.</i></p>
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<h2>I.Calibrations</h2>
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<div class="inlabdiv">
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<p>OD<sub>600</sub> reference point:</p>
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<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>
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<font size = "1">Table 1. OD600 Reference Point.</font>
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</div>
  
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<div class="inlabdiv">
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                                <p>Particle Standard Curve</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: 50%"><br>
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<font size = "1">Figure 1. Particle Standard Curve.</font>
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</div>
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<div class="inlabdiv">
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                                <p>Particle Standard Curve(log scale)</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: 50%"><br>
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<font size = "1">Figure 1. Particle Standard Curve(log scale).</font>
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</div>
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                        <div class="inlabdiv">
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                                <p>Fluorescein Standard Curve</p>
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<img src = "" alt = "Fluorescein Standard Curve" style = "width: 50%"><br>
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<font size = "1">Figure 2. Fluorescein Standard Curve.</font>
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</div>
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                        <div class="inlabdiv">
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                                <p>Fluorescein Standard Curve(log scale)</p>
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<img src = "" alt = "Fluorescein Standard Curve(log scale)" style = "width: 50%"><br>
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<font size = "1">Figure 2. Fluorescein Standard Curve(log scale).</font>
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</div>
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<h2>II.Cell Measurements</h2>
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<div class="inlabdiv">
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<p><b>Fluorescence Raw</b></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: 70%"><br>
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<font size = "1">Table 2. Raw Plate Reader Measurements of Fluorescence Raw at 0 hour.</font>
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</div>
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<div class = "inlabdiv">
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<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>
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</div>
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<div class = "inlabdiv">
 +
<p><b>Abs<sub>600</sub> Raw</b></p>
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<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>
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<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>
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 +
<div>
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<h1>Discussion</h1>
 +
<p></p>
 +
<p></p>
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</div>
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<div>
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<h1>Our Thoughts and Feelings</h1>
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<p></p>
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</div>
  
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</body>
  
  
  
 
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Revision as of 07:12, 5 October 2018

BJRS

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

  1. Negative Control: BBa_R0040
  2. Positive Control: BBa_I20270
  3. Test Device 1: BBa_J364000
  4. Test Device 2: BBa_J364001
  5. Test Device 3: BBa_J364002
  6. Test Device 4: BBa_J364007
  7. Test Device 5: BBa_J364008
  8. 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 Protocol
Help: Protocols/Transformation

Experiment Process

Open Distribution Kit

Add silica beads to 96 well plate

Results

The results are shown in the tables and figures.

I.Calibrations

OD600 reference point:

OD600 Reference Point
Table 1. OD600 Reference Point.

Particle Standard Curve

Particle Standard Curve
Figure 1. Particle Standard Curve.

Particle Standard Curve(log scale)

Particle Standard Curve(log scale)
Figure 1. Particle Standard Curve(log scale).

Fluorescein Standard Curve

Fluorescein Standard Curve
Figure 2. Fluorescein Standard Curve.

Fluorescein Standard Curve(log scale)

Fluorescein Standard Curve(log scale)
Figure 2. Fluorescein Standard Curve(log scale).

II.Cell Measurements

Fluorescence Raw

Fluorescence at 0h
Table 2. Raw Plate Reader Measurements of Fluorescence Raw at 0 hour.
Fluorescence at 6h
Table 3. Raw Plate Reader Measurements of Fluorescence Raw at 6 hours.

Abs600 Raw

Abs600 at 0h
Table 4. Raw Plate Reader Measurements of Abs600 Raw at 0 hour.
Abs600 at 6h
Table 5. Raw Plate Reader Measurements of Abs600 Raw at 6 hours.

Discussion

Our Thoughts and Feelings