Difference between revisions of "Team:Edinburgh UG/Plate Reader"
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| − | + | <li><a href="https://2016.igem.org/Team:Edinburgh_UG/Lexicon_Encoding">Lexicon Encoding</a></li> | |
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| − | + | <li> | |
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| − | + | <li><a href="https://2016.igem.org/Team:Edinburgh_UG/Entrepreneurship">Entrepreneurship</a></li> | |
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| − | + | <li class="dropdown"> | |
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| − | + | <ul class="dropdown-menu" role="menu"> | |
| − | + | <li><a href="https://2016.igem.org/Team:Edinburgh_UG/Plate_Reader">Plate Reader</a></li> | |
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| − | + | <h2 class="section-heading">Introduction</h2> | |
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| + | <p>The aim of the 2016 iGEM InterLab was to quantify the expression of GFP under different promoters and Ribosomal Binding Regions (RBRs). The use of GFP to indirectly measure the expression of a specific protein in cells is a key approach in biology. It enables us to monitor expression levels without killing the cells. The ability measure expression levels per cell is important to be able to quantify strength of promotors. In order to achieve this, population size is indirectly measured using OD600. Expression per cell is then obtained by Fluorescence/OD600.</p> | ||
| + | <br> | ||
| + | <p>Both Fluorescence and OD600 are relative measurements, meaning the instruments used and their settings play a crucial role.</p> | ||
| + | <p>To calibrate the instrument and transform relative measurements into absolutes ones, FITC is used as reference material to construct a standard calibration curve for fluorescence. FITC standard is constructed by serial dilution. The resulting FITC curve is used in combination with another FITC standard curve and a GFP standard curve provided by iGEM.</p> | ||
| + | <p>OD600 is a measure of light scattering, therefore light detection is dependent on the physical geometry of the machine. Calibration experiments using a flow cytometer do not involve standard curves. In order to give uniformity to Fluorescence/OD600 data, a standard scattering solution of a mono-dispersed silica suspension (LUDOX) is used.</p> | ||
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| + | <section id="about"> | ||
| + | <div class="container"> | ||
| + | <div class="row"> | ||
| + | <div class="col-lg-12 text-center"> | ||
| + | <h2 class="section-heading">Plate Reader</h2> | ||
| + | <div class="panel-body"> | ||
| + | <p> All other protocols used in the process of the study can be found on the <a href="https://2016.igem.org/Team:Edinburgh_UG">protocols</a> page.</p> | ||
| + | <p>The aim of the 2016 iGEM InterLab was to quantify the expression of GFP under different promoters and Ribosomal Binding Regions (RBRs). The use of GFP to indirectly measure the expression of a specific protein in cells is a key approach in biology. It enables us to monitor expression levels without killing the cells. The ability measure expression levels per cell is important to be able to quantify strength of promotors. In order to achieve this, population size is indirectly measured using OD600. Expression per cell is then obtained by Fluorescence/OD600.</p> | ||
| + | <br> | ||
| + | <p>Both Fluorescence and OD600 are relative measurements, meaning the instruments used and their settings play a crucial role.</p> | ||
| + | <p>To calibrate the instrument and transform relative measurements into absolutes ones, FITC is used as reference material to construct a standard calibration curve for fluorescence. FITC standard is constructed by serial dilution. The resulting FITC curve is used in combination with another FITC standard curve and a GFP standard curve provided by iGEM.</p> | ||
| + | <p>OD600 is a measure of light scattering, therefore light detection is dependent on the physical geometry of the machine. Calibration experiments using a flow cytometer do not involve standard curves. In order to give uniformity to Fluorescence/OD600 data, a standard scattering solution of a mono-dispersed silica suspension (LUDOX) is used.</p> | ||
| + | </div> | ||
| + | </div> | ||
| + | </div> | ||
| + | <div align="center"> | ||
| + | <a href="#accordion"> | ||
| + | <img src="https://static.igem.org/mediawiki/2016/6/69/Rsz_1arrow.jpg" class="arrow"> | ||
| + | </a> | ||
| + | </div> | ||
| + | <br> | ||
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| + | </div> | ||
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Revision as of 14:31, 14 October 2016
Team:Edinburgh/Part Collection
InterLab Study
Introduction
The aim of the 2016 iGEM InterLab was to quantify the expression of GFP under different promoters and Ribosomal Binding Regions (RBRs). The use of GFP to indirectly measure the expression of a specific protein in cells is a key approach in biology. It enables us to monitor expression levels without killing the cells. The ability measure expression levels per cell is important to be able to quantify strength of promotors. In order to achieve this, population size is indirectly measured using OD600. Expression per cell is then obtained by Fluorescence/OD600.
Both Fluorescence and OD600 are relative measurements, meaning the instruments used and their settings play a crucial role.
To calibrate the instrument and transform relative measurements into absolutes ones, FITC is used as reference material to construct a standard calibration curve for fluorescence. FITC standard is constructed by serial dilution. The resulting FITC curve is used in combination with another FITC standard curve and a GFP standard curve provided by iGEM.
OD600 is a measure of light scattering, therefore light detection is dependent on the physical geometry of the machine. Calibration experiments using a flow cytometer do not involve standard curves. In order to give uniformity to Fluorescence/OD600 data, a standard scattering solution of a mono-dispersed silica suspension (LUDOX) is used.
Plate Reader
All other protocols used in the process of the study can be found on the protocols page.
The aim of the 2016 iGEM InterLab was to quantify the expression of GFP under different promoters and Ribosomal Binding Regions (RBRs). The use of GFP to indirectly measure the expression of a specific protein in cells is a key approach in biology. It enables us to monitor expression levels without killing the cells. The ability measure expression levels per cell is important to be able to quantify strength of promotors. In order to achieve this, population size is indirectly measured using OD600. Expression per cell is then obtained by Fluorescence/OD600.
Both Fluorescence and OD600 are relative measurements, meaning the instruments used and their settings play a crucial role.
To calibrate the instrument and transform relative measurements into absolutes ones, FITC is used as reference material to construct a standard calibration curve for fluorescence. FITC standard is constructed by serial dilution. The resulting FITC curve is used in combination with another FITC standard curve and a GFP standard curve provided by iGEM.
OD600 is a measure of light scattering, therefore light detection is dependent on the physical geometry of the machine. Calibration experiments using a flow cytometer do not involve standard curves. In order to give uniformity to Fluorescence/OD600 data, a standard scattering solution of a mono-dispersed silica suspension (LUDOX) is used.
