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Team:Groningen/flowcytometry
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=Flowcytometry protocol= | =Flowcytometry protocol= | ||
<br> | <br> | ||
| - | <br> Inoculate 10ml M9 minimal medium + 1ml of 2% casamino acids and incubate overnight at 37 °C | + | <br> Inoculate 10ml M9 minimal medium + antibiotics + 1ml of 2% casamino acids and incubate overnight at 37 °C. |
| - | + | <br> The next day, the pre-heated erlenmeyers containing the 10ml M9 minimal medium+ antibiotics + 1ml of 2% casamino <br> acids of are inoculated until an OD600 of 0.01. | |
| - | <br> The next day, the pre-heated erlenmeyers containing the 10ml M9 minimal medium | + | |
<br> The FACS can be prepared for measurements with the protocol provided by Maarten Mols: | <br> The FACS can be prepared for measurements with the protocol provided by Maarten Mols: | ||
<br> [[File:Flowcytometryprotocol.pdf]] | <br> [[File:Flowcytometryprotocol.pdf]] | ||
<br> | <br> | ||
| - | <br> 100μl suspension of the logfase cells will be used for measurements with the FACS | + | <br> 100μl suspension of the logfase cells will be used for measurements with the FACS. |
| - | <br> Later on, when the celldensity increases too much, | + | <br> Later on, when the celldensity increases too much, the cells will be diluted with M9 media. |
<br> | <br> | ||
<br> The PBADaraC promotor needs to be induced by arabinose. | <br> The PBADaraC promotor needs to be induced by arabinose. | ||
Revision as of 15:05, 21 September 2011
Flowcytometry
In this project, we wanted to measure promotor activities and their leakage, but also testing the constructs
whether they work or not. Analysis of these constructs was done with the help of the FACS for measuring
fluorescence.
Flowcytometry protocol
Inoculate 10ml M9 minimal medium + antibiotics + 1ml of 2% casamino acids and incubate overnight at 37 °C.
The next day, the pre-heated erlenmeyers containing the 10ml M9 minimal medium+ antibiotics + 1ml of 2% casamino
acids of are inoculated until an OD600 of 0.01.
The FACS can be prepared for measurements with the protocol provided by Maarten Mols:
File:Flowcytometryprotocol.pdf
100μl suspension of the logfase cells will be used for measurements with the FACS.
Later on, when the celldensity increases too much, the cells will be diluted with M9 media.
The PBADaraC promotor needs to be induced by arabinose.
Preparation for 10% arabinose stock:
Add 1g of arabinose to 9ml milliQ and filter sterilize the stock.
Concentrations for induction can be varied from 0 to 1%.
See for more information: http://openwetware.org/wiki/Arabinose
