Penn State University 2006:Plate-Reading: Difference between revisions
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This is a kinetic assay for induction by HSL over a 107-magnitude concentration. Each construct is measured in triplicate. | This is a kinetic assay for induction by HSL over a 107-magnitude concentration. Each construct is measured in triplicate. | ||
===<font color="red">Media/Reagents/Materials</font>=== | |||
Plates that are needed: 3+X shaker/growing, X fluorescence, where 2X is number of time points desired. | Plates that are needed: 3+X shaker/growing, X fluorescence, where 2X is number of time points desired. | ||
*M9+0.1%YE+antibiotic plates | |||
*210 mL M9+0.1%YE+antibiotic | |||
*96 well growing plates | |||
*96 well fluorescence plates | |||
*96 well clear bottom absorbance plates | |||
===<font color=" | ===<font color="orange">Pre</font>=== | ||
M9+0.1%YE+antibiotic plates | #Condition parts on M9+0.1%YE+antibiotic minimal media plates | ||
#Inoculate 3 single colonies from these plates into M9+0.1%YE+antibiotic media overnight | |||
96 well growing plates | #Autoclave 96 well shaker/growing plates | ||
#Dilute cells down to OD600=0.1 | |||
===<font color="blue">Plate Reading</font>=== | |||
#Do not put cells on ice! | |||
Plate Reading | |||
For GFP, assay at 30ºC | For GFP, assay at 30ºC | ||
'' | '' | ||
Revision as of 23:04, 24 October 2006
Plate Reading
Plate Reading
Time: variable
About
This is a kinetic assay for induction by HSL over a 107-magnitude concentration. Each construct is measured in triplicate.
Media/Reagents/Materials
Plates that are needed: 3+X shaker/growing, X fluorescence, where 2X is number of time points desired.
- M9+0.1%YE+antibiotic plates
- 210 mL M9+0.1%YE+antibiotic
- 96 well growing plates
- 96 well fluorescence plates
- 96 well clear bottom absorbance plates
Pre
- Condition parts on M9+0.1%YE+antibiotic minimal media plates
- Inoculate 3 single colonies from these plates into M9+0.1%YE+antibiotic media overnight
- Autoclave 96 well shaker/growing plates
- Dilute cells down to OD600=0.1
Plate Reading
- Do not put cells on ice!
For GFP, assay at 30ºC