Difference between revisions of "Team:NAWI Graz/InterLab"
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| − | + | Reliable and repeatable measurement is key to compare and analyse data from different labs all over the world. To support the effort to create detailed protocols to measure fluorescence and improve the possibility of comparing data, the iGEM Team NAWI_Graz 2017 decided to participate in the <a href=”https://2017.igem.org/Competition/InterLab_Study”>Fourth International InterLaboratory Measurement Study</a> in synthetic biology. The goal is to establish a GFP measurement protocol based on engineering principles. | |
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| − | + | Our tasks were to follow exactly the <a href=”https://static.igem.org/mediawiki/2017/8/85/InterLab_2017_Plate_Reader_Protocol.pdf”>iGEM Plate Reader Protocol< /a>, fill in the prepared excel sheets and send the results to the iGEM headquarter. | |
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<h2 class="section-sub">Results</h2> | <h2 class="section-sub">Results</h2> | ||
<div class="col section-text container"> | <div class="col section-text container"> | ||
| − | + | For this study we needed to transform 8 devices into the presupposed bacteria strain <i>E.coli</i> DH5 alpha. This included a Positive Control <a href=”http://parts.igem.org/Part:BBa_I20270”>I20270< /a>, the constitutive TetR repressible promoter as Negativ Control <a href=”http://parts.igem.org/Part:BBa_R0040”>R0040< /a>, Test Device 1 <a href=”http://parts.igem.org/Part:BBa_J364000”>J364000< /a>, Test Device 2 <a href=”http://parts.igem.org/Part:BBa_J364001”>J364001< /a>, Test Device 3 <a href=”http://parts.igem.org/Part:BBa_J364002”>J364002< /a>, Test Device 4 <a href=”http://parts.igem.org/Part:BBa_J364003”>J364003< /a>, Test Device 5 <a href=”http://parts.igem.org/Part:BBa_J364004”>J364004< /a> and Test Device 6 <a href=”http://parts.igem.org/Part:BBa_J364005”>J364005< /a>. They are all stored in <a href=”http://parts.igem.org/Part:pSB1C3”>pSB1C3< /a> and were cultured on LB-Agar plates with a chloramphenicol concentration of 100 µg/ml. As we had grown colonies the day after the transformations, we could start with the cell measurement protocols. | |
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<h2 class="section-sub">Calibration Protocols</h2> | <h2 class="section-sub">Calibration Protocols</h2> | ||
| − | <h3 class="section-sub-sub"> | + | <h3 class="section-sub-sub">1. OD<sub>600</sub> Reference point</h3> |
<div class="section-sub-text container"> | <div class="section-sub-text container"> | ||
| − | + | Results of OD<sub>600</sub> measurement of 1 ml LUDOX and 1 ml H<sub>2</sub>O with dH<sub>2</sub>O as blank. | |
</div> | </div> | ||
<div class="col section-image border border-secondary"> | <div class="col section-image border border-secondary"> | ||
<img src="" alt="[protocols table]"> | <img src="" alt="[protocols table]"> | ||
| + | Tab. 1: OD<sub>600</sub> measurement | ||
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<h3 class="section-sub-sub">Protocol Fluorescein Fluorescence Standard Curve</h3> | <h3 class="section-sub-sub">Protocol Fluorescein Fluorescence Standard Curve</h3> | ||
Revision as of 16:38, 31 October 2017
INTERLAB MEASUREMENT STUDY
Reliable and repeatable measurement is key to compare and analyse data from different labs all over the world. To support the effort to create detailed protocols to measure fluorescence and improve the possibility of comparing data, the iGEM Team NAWI_Graz 2017 decided to participate in the Fourth International InterLaboratory Measurement Study in synthetic biology. The goal is to establish a GFP measurement protocol based on engineering principles.
Our tasks were to follow exactly the iGEM Plate Reader Protocol< /a>, fill in the prepared excel sheets and send the results to the iGEM headquarter.
Results
For this study we needed to transform 8 devices into the presupposed bacteria strain E.coli DH5 alpha. This included a Positive Control I20270< /a>, the constitutive TetR repressible promoter as Negativ Control R0040< /a>, Test Device 1 J364000< /a>, Test Device 2 J364001< /a>, Test Device 3 J364002< /a>, Test Device 4 J364003< /a>, Test Device 5 J364004< /a> and Test Device 6 J364005< /a>. They are all stored in pSB1C3< /a> and were cultured on LB-Agar plates with a chloramphenicol concentration of 100 µg/ml. As we had grown colonies the day after the transformations, we could start with the cell measurement protocols.
Calibration Protocols
1. OD600 Reference point
Results of OD600 measurement of 1 ml LUDOX and 1 ml H2O with dH2O as blank.
Protocol Fluorescein Fluorescence Standard Curve
We measured the in PBS diluted fluorescein 96 well plate in a plate reader with the settings XY and turned of pathlength
correction.
Cell Measurement Protocol
- Day 1 OD600 Measurement
- Day 2 Transformation
- Day 3 Overnight culture
InterLab
Bronze Medal Criterion #4
Standard Tracks: Participate in the Interlab Measurement Study and/or improve the characterization of 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 2017 part number range.
For teams participating in the InterLab study, all work must be shown on this page.
