Microplate Reader Quick Start

  1. It's always best to start from single colony picks that had been grown overnight in liquid culture to saturation.
  2. A typical start culture dilution is 2:200, 1:200 or 1:250. The most diluted one (1:250) would provide the best result for measuring lag phase times.
  3. Plan out which wells will contain what sample. Randomizing a bit can help average out temperature differences throughout the plate; you can use a simple "JumpTwoRowsDown-Jump#ofReplicatesToRight" algorithm for positioning each replicate in the 96-well plate. Printable 96 well plate templates are available online. Generally, each condition should be done in quadruplicate (or triplicate depending on space concerns). DON'T FORGET TO INCLUDE MEDIA BLANKS (minimum of one!)
  4. Sign up and make reservation for TECAN microplate reader here TECAN microplate
  5. Select the appropriate type of plate for your application: black walled, clear bottom for fluorescence (white walled, clear bottom is generally for luminescence and completely clear is generally for other applications, like measuring OD600).
  6. Ensure that you have adequate controls for your experiment. At the very least, have your negative control in the same media grown under the same conditions.
  7. Turn on the plate reader - there is a power button in the back next to the power cord - and then turn on the Magellan software on the computer next to it. Not doing it in this order may cause the software to fail to recognize the machine.
  8. Select the option that says "raw data" and eventually you will find yourself at the configuration screen.
  9. Once the protocol is started an Excel worksheet will open up automatically to record the data.

For OD600 measurements and growth rates go to Determining Growth Rates

For fluorescence measurements go to Fluorescence Measurements

--Main.GabrielSuarez - 14 Dec 2017

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Contributors to this topic Edit topic GabrielSuarez
Topic revision: r2 - 2017-12-27 - 02:06:34 - Main.GabrielSuarez
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