IGEM:IMPERIAL/2007/Projects/Hrp System/Systems/Hrp Device 1/TestingValidation/Device1protocol

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Contents

Equipment

  • Twinkle Plate Reader Fluorometer
  • 96 well plate
  • Eppendorf Tubes
  • Gilson Pippettes
  • Plastic Curvetts
  • Incubator 37o shaker
  • Vortex and Centrifuge
  • Spectrometer

Materials and Reagents

  • M9 minimal media 50μg/ml Ampicillin
  • Inducer of promoter at varying concentrations
  • Recombinant acGFP of known concentration
  • Agar Plates
  • E.coli strain containing vector

Protocols

Promoter Test Construct

The promoter pLux is to be tested at 8 varying ranges of inducer concentration and each conbcentration will haev 4 repeats.

  • pLux will have a range of 0 to 100μM AHL added at intervals of 12.5μM Protocol

Device Test Construct

For this test construct we initially need to run a preliminary test in order to find out the time intervals and input ranges we need. We will start the preliminary test using the range of concentrations used initially for promoters. This device uses a pLac promoter, the proposed range of concentrations is going to be defined from the promoter experiment, this is use a range of concentrations to cover an appropriate range of promoter. From looking at literature an appropriate range is 0-100μM.

  • First the is carried out. From this the device 1 protocol may wished to be changed. Click here for the preliminary protocol.
  • Once the data range has been adjusted then carry out the test construct protocol. Click here for the device protocol
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