Difference between revisions of "Team:SDU-CHINA/InterLab"

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<h1 class="main-title">Interlab</h1>
 
<h1 class="main-title">Interlab</h1>
 
<div class="paragraghs">
 
<div class="paragraghs">
<p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;The InterLab protocol aims to address these issues by providing researchers with a detailed protocol and data analysis form that yields absolute units for measurements of GFP in a plate reader. We were invited to participate in the Fifth International Interlaboratory Measurement Study. The goal of the iGEM InterLab Study is to identify and correct the sources of
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<p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;The InterLab protocol aims to address key issues in synthetic biology by providing researchers with detailed protocol and data analysis. In our case, we measure the absolute units for measurements of GFP in a plate reader. We were invited to participate in the Fifth International Interlaboratory Measurement Study. The goal of the iGEM InterLab Study is to identify and correct the sources of
 
systematic variability in synthetic biology measurements, so that eventually, measurements that are taken in different labs will be no more variable than measurements taken within the
 
systematic variability in synthetic biology measurements, so that eventually, measurements that are taken in different labs will be no more variable than measurements taken within the
 
same lab. In order to compute the cell count in our samples, we use two orthogonal approaches:</p>
 
same lab. In order to compute the cell count in our samples, we use two orthogonal approaches:</p>
 
<ol>
 
<ol>
<li>Converting between absorbance of cells to absorbance of a known concentration of beads.</li>
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<li>Converting between absorbance of cells and absorbance of a known concentration of beads.</li>
 
<li>Counting colony-forming units (CFUs) from the sample.</li>
 
<li>Counting colony-forming units (CFUs) from the sample.</li>
 
</ol>
 
</ol>
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measurements.</p>
 
measurements.</p>
 
<br />
 
<br />
<p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;We followed the protocol provided by committee and finished all the measurement during July.</p>
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<p>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;We followed the protocol provided by the committee and finished all the measurement during July.</p>
 
<p><b>Day 1</b>​: We successfully transformed ​Escherichia coli​ DH5α with following plasmids:</p>
 
<p><b>Day 1</b>​: We successfully transformed ​Escherichia coli​ DH5α with following plasmids:</p>
 
<table>
 
<table>
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</tr>
 
</tr>
 
</table>
 
</table>
<p><b>Day 2​</b>: We picked 2 colonies respectively from every transformation plates and inoculated in 5-10 mL LB medium +Chloramphenicol. Then the cells were cultured overnight (16-18 hours) at 37°C and 220 rpm.</p>
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<p><b>Day 2​</b>: We picked 2 colonies respectively from every transformation plates and inoculated in 5-10 mL LB medium + Chloramphenicol. Then the cells were cultured overnight (16-18 hours) at 37°C and 220 rpm.</p>
 
<p><b>Day 3</b>: Firstly, we took three calibration measurements: an OD ​ 600 reference point, a particle standard curve, and a fluorescein standard curve.</p>
 
<p><b>Day 3</b>: Firstly, we took three calibration measurements: an OD ​ 600 reference point, a particle standard curve, and a fluorescein standard curve.</p>
 
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Revision as of 05:56, 30 September 2018

Interlab

      The InterLab protocol aims to address key issues in synthetic biology by providing researchers with detailed protocol and data analysis. In our case, we measure the absolute units for measurements of GFP in a plate reader. We were invited to participate in the Fifth International Interlaboratory Measurement Study. The goal of the iGEM InterLab Study is to identify and correct the sources of systematic variability in synthetic biology measurements, so that eventually, measurements that are taken in different labs will be no more variable than measurements taken within the same lab. In order to compute the cell count in our samples, we use two orthogonal approaches:

  1. Converting between absorbance of cells and absorbance of a known concentration of beads.
  2. Counting colony-forming units (CFUs) from the sample.

      By using these two approaches, we will be able to determine how much they agree with each other, and whether using one (or both) can help to reduce lab-to-lab variability in measurements.


      We followed the protocol provided by the committee and finished all the measurement during July.

Day 1​: We successfully transformed ​Escherichia coli​ DH5α with following plasmids:

Device Part Number Plate Location
Negative control BBa_R0040 Kit Plate 7 Well 2D
Positive control BBa_I20270 Kit Plate 7 Well 2B
Test Device 1 BBa_J364000 Kit Plate 7 Well 2F
Test Device 2 BBa_364001 Kit Plate 7 Well 2H
Test Device 3 BBa_364002 Kit Plate 7 Well 2J
Test Device 4 BBa_364007 Kit Plate 7 Well 2L
Test Device 5 BBa_364008 Kit Plate 7 Well 2N
Test Device 6 BBa_364008 Kit Plate 7 Well 2P

Day 2​: We picked 2 colonies respectively from every transformation plates and inoculated in 5-10 mL LB medium + Chloramphenicol. Then the cells were cultured overnight (16-18 hours) at 37°C and 220 rpm.

Day 3: Firstly, we took three calibration measurements: an OD ​ 600 reference point, a particle standard curve, and a fluorescein standard curve.

Secondly, we did cell measurement. From 0 hour to 6 hour, we did CFU measurement and coated all 36 plates. After that, we measured Abs600 and fluorescence of the cultures in the 96 wells plate in the plate reader.

Day 4: We counted the colonies on every plate with fewer than 300 colonies. Our experimental data can be seen here.

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