Revision as of 19:19, 19 October 2016

INTERLAB STUDY

Building up on last years findings (Beal, Jacob, et al.¹), the Third Annual Interlaboratory Measurement Study was continued during this years iGEM competition. The main focus still lays on the comparison of fluorescence measurements of identical constructs from different labs and with different equipment. This years question to explore: "How close can the numbers be when fluorescence is measured all around the world?"

PROTOCOL

This years plate reader protocol was straight-forward. The protocol to measure all devices using flow cytometry however, was not yet up to the same standard. To make participating in next years InterLab study a bit more attractive, we propose an easy-to-use FACS protocol as well as a new Excel sheet for data collection:

Figure 1: New cell measurement protocol for flow cytometry analysis of the devices. We recommend these improvements to the existing protocol, since it grants a better overview of the current and following steps, thereby decreasing possible sources of errors.

RESULTS

Figure 2: Device 2 (J23106+I13504) compared to negative control. As expected, fluorescence intensity of Device 2 increases over time.

We measured all five devices with our plate reader (Tecan Infinite m200PRO) as well as our departments flow cytometer (BD LSRFortessa). The submitted data can be found here: Plate reader data or flow cytometry data. The following devices were tested:

Test Device 1: J23101.B0034.E0040.B0015 in pSB1C3
Test Device 2: J23106.B0034.E0040.B0015 in pSB1C3
Test Device 3: J23117.B0034.E0040.B0015 in pSB1C3
Positive Control Device: I20270 in pSB1C3
Negative Control Device: R0040 in pSB1C3

CONCLUSION

Participating in this years InterLab Study was feasible in the given timescale. However, we feel like the data submission deadline should be moved closer towards end of September, since not every team has access to their laboratory space early enough to get comfortable with the equipment and procedures. This could potentially increase the submitted data quality. We highly recommend teams of future iGEM competitors to sign up and we are looking forward to the resulting comparison of the measured data.

REFERENCES

[1] Beal, Jacob, et al. "Reproducibility of Fluorescent Expression from Engineered Biological Constructs in E. coli." PloS one 11.3 (2016): e0150182.

@@ Line 27: / Line 27: @@
-<div class="image full_size">
+<div class="image_box full_size">
      <a href="https://2016.igem.org/File:T--ETH_Zurich--Protocol_Flow_Cytometry.pdf">
          <img src="https://static.igem.org/mediawiki/2016/b/b0/T--ETH_Zurich--Updated_Facs_Protocol_2sided.jpg">
+        <p><b>Figure 1:</b> New cell measurement protocol for flow cytometry analysis of the devices. We recommend these improvements to the existing protocol, since it grants a better overview of the current and
+        following steps, thereby decreasing possible sources of errors.
+        </p>
      </a>
 </div>
@@ Line 47: / Line 50: @@
    <h2>RESULTS</h2>
    <p>
+<div class="image_box">
-We measured all five devices with our plate reader as well as the departments flow cytometer. The submitted data can be found here: <a href="https://static.igem.org/mediawiki/2016/b/b0/T--ETH_Zurich--plate_reader_data.xls"> Plate reader data</a> or <a href="https://static.igem.org/mediawiki/2016/1/1c/T--ETH_Zurich--flow_cytometry_data.xlsx">flow cytometry data</a>. An example graph comparing Device 2 (J23106+I13504) to the negative control is shown here:
-<div class="image_box" style="max-width: 3000px;">
      <a href="https://static.igem.org/mediawiki/2016/f/f4/T--ETH_Zurich--Example_Plot_InterLab1.png">
          <img src="https://static.igem.org/mediawiki/2016/f/f4/T--ETH_Zurich--Example_Plot_InterLab1.png">
+        <p><b>Figure 2: </b>Device 2 (J23106+I13504) compared to negative control. As expected, fluorescence intensity of Device 2 increases over time.</p>
      </a>
 </div>
+We measured all five devices with our plate reader (Tecan Infinite m200PRO) as well as our departments flow cytometer (BD LSRFortessa). The submitted data can be found here: <a href="https://static.igem.org/mediawiki/2016/b/b0/T--ETH_Zurich--plate_reader_data.xls"> Plate reader data</a> or <a href="https://static.igem.org/mediawiki/2016/1/1c/T--ETH_Zurich--flow_cytometry_data.xlsx">flow cytometry data</a>. The following devices were tested:<br>
+<ul><li>Test Device 1: J23101.B0034.E0040.B0015 in pSB1C3</li>
+    <li>Test Device 2: J23106.B0034.E0040.B0015 in pSB1C3</li>
+    <li>Test Device 3: J23117.B0034.E0040.B0015 in pSB1C3</li>
+    <li>Positive Control Device: I20270 in pSB1C3</li>
+    <li>Negative Control Device: R0040 in pSB1C3</li>
+</ul>
@@ Line 67: / Line 78: @@
-Participating in this years InterLab Study did not take too much effort. However, we feel like the data submission deadline should be moved closer towards end of
+Participating in this years InterLab Study was feasible in the given timescale. However, we feel like the data submission deadline should be moved closer towards end of
-September, since not every team has access to their laboratory space early enough to get comfortable with the equipment and procedures. We highly recommend teams of future
+September, since not every team has access to their laboratory space early enough to get comfortable with the equipment and procedures. This could potentially increase the submitted data quality.
-iGEM competitors to sign up and we are looking forward to the resulting comparison of the measured data.
+We highly recommend teams of future iGEM competitors to sign up and we are looking forward to the resulting comparison of the measured data.

Difference between revisions of "Team:ETH Zurich/InterLab"