Difference between revisions of "Team:Toronto/InterLab"
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<p>Table shows absorbance measurements (at 600 nm) for LUDOX CL-X and dd-H2O using a plate reader. The corrected Abs<sub>600</sub> is the difference between the LUDOX CL-X reading and dd H2O reading. Reference OD<sub>600</sub> is a measurement by a spectrophotometer (provided on iGEM excel sheet). OD<sub>600</sub>/Abs<sub>600</sub> is the correction factor to convert Abs<sub>600</sub> to OD<sub>600</sub>, calculated by dividing Reference OD<sub>600</sub> by Abs<sub>600</sub>.</p> | <p>Table shows absorbance measurements (at 600 nm) for LUDOX CL-X and dd-H2O using a plate reader. The corrected Abs<sub>600</sub> is the difference between the LUDOX CL-X reading and dd H2O reading. Reference OD<sub>600</sub> is a measurement by a spectrophotometer (provided on iGEM excel sheet). OD<sub>600</sub>/Abs<sub>600</sub> is the correction factor to convert Abs<sub>600</sub> to OD<sub>600</sub>, calculated by dividing Reference OD<sub>600</sub> by Abs<sub>600</sub>.</p> | ||
<img src="https://static.igem.org/mediawiki/2018/d/d0/T--Toronto--7_25_2018_Interlab_LUDOX_CL_X.png" alt="LUDOX_CL_X"> | <img src="https://static.igem.org/mediawiki/2018/d/d0/T--Toronto--7_25_2018_Interlab_LUDOX_CL_X.png" alt="LUDOX_CL_X"> | ||
Revision as of 00:58, 26 July 2018
- Purpose of this calibration: To transform absorbance data to a OD600 measurement, calculate a plate-reader specific (Tecan Infinite 200 Pro) conversion factor for OD600 from Abs600 calculated for Ludox CL-X on a mass spectrophotometer.
- Beer-Lambert’s law of absorbance dictates that optical path length plays a fundamental role in determining absorbance:
- This is necessary because cuvettes (used in photometer) have a fixed path optical path length when using light scattering to measure absorbance as opposed to the varying path lengths of wells in a 96-well plate which change as the volume of sample added in them changes.
- Results: Cell density readings can thus be converted to OD600 by multiplying correction factor value, 4.138.
Table1.
Table shows absorbance measurements (at 600 nm) for LUDOX CL-X and dd-H2O using a plate reader. The corrected Abs600 is the difference between the LUDOX CL-X reading and dd H2O reading. Reference OD600 is a measurement by a spectrophotometer (provided on iGEM excel sheet). OD600/Abs600 is the correction factor to convert Abs600 to OD600, calculated by dividing Reference OD600 by Abs600.
