(r1) MicroplateReaderQuickStartGuide < Lab

---+ Microplate Reader Quick Start

   1 It's always best to start from single colony picks that had been preconditioned overnight in liquid culture to saturation.
   1 Typical dilution for OD600 measurements is 2:200, 1:200 or 1:250, since the latter is most diluted it would provide the best result for measuring lag phase.
   1 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 <span style="background-color: transparent">[[http://www.cellsignet.com/media/plates/96.jpg][online]]</span><span style="background-color: transparent">. Generally, each condition should be done in quadruplicate (or triplicate depending on space concerns). DON'T FORGET TO INCLUDE MEDIA BLANKS (minimum of one!)</span>
   1 Sign up and make reservation for TECAN microplate reader here [[https://tecanplatereader.skedda.com/booking][TECAN microplate]]
   1 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).
   1 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.
   1 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.
   1 Select the option that says "raw data" and eventually you will find yourself at the configuration screen.
   1 Once the protocol is started an Excel worksheet will open up automatically to record the data.

For *OD600 measurements* and *growth rates* go to [[http://barricklab.org/twiki/bin/view/Lab/ProtocolsGrowthRates][Determining Growth Rates]]

For *fluorescence* *measurements* go to [[http://barricklab.org/twiki/bin/view/Lab/ProtocolsFluorescentProteinEvolutionaryStability][Fluorescence Measurements]]

--Main.GabrielSuarez - 14 Dec 2017
Barrick Lab > ProtocolList > MicroplateReaderQuickStartGuide
Topic revision: r1 - 2017-12-14 - 22:17:38 - Main.GabrielSuarez