Magni:Protocols/tecan: Difference between revisions
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(New page: {{Magni}} <div style="padding: 10px; width: 720px; border: 5px solid #000000;"> = Infinite F200 readings = The microplate reader can be used for static or dynamic readings of bacteria cul...) |
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* Dispense deionized water in all the unused wells. | * Dispense deionized water in all the unused wells. | ||
* Add inducers to the cultures when required. | * Add inducers to the cultures when required. | ||
* Start a kinetic cycle of 18 hours with the i-control software (Tecan): 37degC during all the experiment, linear shaking (15 s, 3 mm amplitude), 5 s wait time (also 10 s is ok), absorbance reading (default parameters), fluorescence readings (gain=50 for medium and high copy plasmids, gain=80 for low copy plasmids, gain=80 or 100 for single-copy integrants), sampling time=5 min. The specified fluorescence gain factors are only guidelines and can be modified according to promoter/RBS strength, strain and cell density. | * Start a kinetic cycle of 18 hours with the i-control software (Tecan): 37degC during all the experiment, linear shaking (15 s, 3 mm amplitude), 5 s wait time (also 10 s is ok), absorbance reading (default parameters), fluorescence readings (gain=50 for medium and high copy plasmids, gain=50 or 80 for low copy plasmids, gain=80 or 100 for single-copy integrants), sampling time=5 min. The specified fluorescence gain factors are only guidelines and can be modified according to promoter/RBS strength, strain, growth medium and cell density. | ||
* Always include a "blank" for absorbance and fluorescence, as described before. | * Always include a "blank" for absorbance and fluorescence, as described before. | ||
Revision as of 10:29, 2 February 2013
Infinite F200 readings
The microplate reader can be used for static or dynamic readings of bacteria cultures growth and fluorescent proteins.
Static readings
- Grow bacteria under the desired conditions.
- Transfer 200 ul of bacteria in a well of a 96-well microplate.
- Measure with or without lid.
- Always measure a "blank" that can be subtracted to the other raw measurements (blank for absorbance is sterile media; blank for absorbance is non-fluorescent culture grown to an OD600 comparable to the fluorescent cultures).
Dynamic readings
- Grow bacteria under the desired conditions.
- Transfer 200 ul of bacteria in a well of a 96-well microplate, excluding the 36 wells of the "frame".
- Dispense deionized water in all the unused wells.
- Add inducers to the cultures when required.
- Start a kinetic cycle of 18 hours with the i-control software (Tecan): 37degC during all the experiment, linear shaking (15 s, 3 mm amplitude), 5 s wait time (also 10 s is ok), absorbance reading (default parameters), fluorescence readings (gain=50 for medium and high copy plasmids, gain=50 or 80 for low copy plasmids, gain=80 or 100 for single-copy integrants), sampling time=5 min. The specified fluorescence gain factors are only guidelines and can be modified according to promoter/RBS strength, strain, growth medium and cell density.
- Always include a "blank" for absorbance and fluorescence, as described before.
