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<img src="https://static.igem.org/mediawiki/2018/b/b5/T--Toronto--7_25_2018_Interlab_lambert-law.gif" alt="lambert-law"> | <img src="https://static.igem.org/mediawiki/2018/b/b5/T--Toronto--7_25_2018_Interlab_lambert-law.gif" alt="lambert-law"> | ||
<li>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.</li> | <li>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.</li> | ||
− | <li>Results: Cell density readings can thus be converted to | + | <li>Results: Cell density readings can thus be converted to OD<sub>600</sub> by multiplying correction factor value, <b>4.138</b>.</li> |
</ul> | </ul> | ||
Revision as of 00:50, 26 July 2018
Calibration 1: OD600 Reference point
- 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.