Difference between revisions of "Team:NTU-Singapore/Measurement"
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<li>Emission Wavelength: 520nm</li> | <li>Emission Wavelength: 520nm</li> | ||
<li>Gain: 40</li> | <li>Gain: 40</li> | ||
+ | <li>Bandwidth: 20</li> | ||
</ol> | </ol> |
Revision as of 15:58, 18 September 2015
Measurement
Fluorescence measurement
Material
- LB
- MQ water
- Micro-centrifuge
- 96-well microplate
- 50ml Falcon Tube
- Tecan Infinite m200 microplate reader
- PBS solution
Procedures
- Inoculate strains for overnight culture in 5mL LB
- Measure the OD600 of the overnight culture with 200uL of sample
- Dilute OD600 to 0.05 in 5mL of LB in 15mL tube
- Incubate at 30C for 4 hours so that bacteria reaches mid-log phase
- Dilute OD600 to 0.05 in 10mL of LB in 50mL tube
- Take 400uL of samples from culture media at one hour intervals starting from T=1hr
- Centrifuge at 15000rpm for 3 minutes
- Discard supernatant and resuspend pellet with 400uL PBS
- Take 150uL of sample and load onto microplate
- Measure OD600 and fluorescence intensity
Tecan Infinite M200 Measurement
Material
Parameters
- Excitation Wavelength: 485nm
- Emission Wavelength: 520nm
- Gain: 40
- Bandwidth: 20
Spotting
Material
- LB agar plate
- Microplate Reader
- LB broth
Parameters
- Grow overnight cultures of sample in 5mL LB broth
- Measure OD600 with 200uL sample
- Dilute to ODOD600 = 0.4
- Spot 3uL of sample onto LB agar plate
- Incubate overnight at 30C
- View with DarkReader Transluminator