Team:Washington/GeneralProtocol

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1. Transform plasmids containing, for example, PLPCB (chromophore) and pJT119B (green light sensor and GFP reporter) into chemically competent JT2 cells.</br>
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1. Transform plasmids containing, for example, PLPCB (chromophore) and pJT119B (green light sensor and GFP reporter) into chemically competent JT2 cells. For full list of plasmids see the Light Sensing Experiments page. </br>
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2. Plate cells on LB agar plates supplemented with the appropriate antibiotics and incubate at 37C overnight. </br>
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2. Plate cells on LB agar plates supplemented with the appropriate antibiotics and incubate at 37C overnight.</br>
See table below for list of systems and plasmids with their required antibiotics.</br>
See table below for list of systems and plasmids with their required antibiotics.</br>
3. Inoculate 5 mL of LB medium with a single colony in a 50 mL Falcon tube.</br>
3. Inoculate 5 mL of LB medium with a single colony in a 50 mL Falcon tube.</br>
4. Grow cultures to saturation overnight at 37C with shaking.</br>
4. Grow cultures to saturation overnight at 37C with shaking.</br>
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5. Determine cell density (OD600 measurement), and dilute overnight cultures to a final OD600 of 1x10E-4 in M9 minimal medium supplemented with casamino acids with the appropriate antibiotics.
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5. Determine cell density using the spectrophotometer(OD600 measurement), and dilute overnight cultures to a final OD600 of 1x10E-4 in M9 minimal medium supplemented with CAS amino acids with the appropriate antibiotics. </br>
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Revision as of 02:56, 27 September 2013



Light Testing General Protocol
1. Transform plasmids containing, for example, PLPCB (chromophore) and pJT119B (green light sensor and GFP reporter) into chemically competent JT2 cells. For full list of plasmids see the Light Sensing Experiments page.
2. Plate cells on LB agar plates supplemented with the appropriate antibiotics and incubate at 37C overnight.
See table below for list of systems and plasmids with their required antibiotics.
3. Inoculate 5 mL of LB medium with a single colony in a 50 mL Falcon tube.
4. Grow cultures to saturation overnight at 37C with shaking.
5. Determine cell density using the spectrophotometer(OD600 measurement), and dilute overnight cultures to a final OD600 of 1x10E-4 in M9 minimal medium supplemented with CAS amino acids with the appropriate antibiotics.

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