Difference between revisions of "Team:Manchester/InterLab"

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Measuring the GFP expression is a useful tool worldwide for quantifying gene expression. Currently, there are various differences in fluorescence data which is measured on different devices (e.g. different units of fluorescence recorded or different settings of the data processing used). The aim of the Fifth InterLab study is to make fluorescence measurements more compatible among various laboratories around the world. This is to be done by standardising the protocol for the machine calibration with Silica beads and relating OD600 values with colony forming units (CFUs). The expression of the GFP by one cell will be calculated by dividing the fluorescence of a population of cells via CFUs instead of using an OD600 measurement which is less precise.
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<p>Measurement of GFP fluorescence is a useful tool for quantifying gene expression. When carrying out this type of work, researchers often wish to state their data as a measurement of fluorescence per cell. Currently, the fluorescence per cell measurement can be variable due to differences between:</p>
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<p>To find fluorescence per cell values, typically the measured fluorescence is divided by the OD<sub>600</sub> as an approximation of a cell number. This can also be imprecise and variable when measured on different devices.  
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<p>The aim of the Fifth InterLab study is to make fluorescence per cell measurements more compatible among various laboratories around the world by investigating the fluorescence of eight test devices built from BioBrick parts. By standardising the protocol for the plate reader calibration with LUDOX solution, silica beads, and fluorescein, we hope to reduce variability between our results and those of other iGEM teams. OD<sub>600</sub> values will be associated with colony forming units (CFUs) to achieve a more accurate measurement of fluorescence per cell. </p>
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Revision as of 18:30, 8 October 2018

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InterLab Study

- The Fifth InterLab Study -

Measurement of GFP fluorescence is a useful tool for quantifying gene expression. When carrying out this type of work, researchers often wish to state their data as a measurement of fluorescence per cell. Currently, the fluorescence per cell measurement can be variable due to differences between:

  • units of measurement
  • data processing settings
  • device model

To find fluorescence per cell values, typically the measured fluorescence is divided by the OD600 as an approximation of a cell number. This can also be imprecise and variable when measured on different devices.

The aim of the Fifth InterLab study is to make fluorescence per cell measurements more compatible among various laboratories around the world by investigating the fluorescence of eight test devices built from BioBrick parts. By standardising the protocol for the plate reader calibration with LUDOX solution, silica beads, and fluorescein, we hope to reduce variability between our results and those of other iGEM teams. OD600 values will be associated with colony forming units (CFUs) to achieve a more accurate measurement of fluorescence per cell.

- Protocols -

All InterLab protocols were provided by iGEM

- Results -