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Team:HokkaidoU Japan/Notebook/aggregation protocols
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| - | + | ==Aggregation Protocols== | |
| - | ==Aggregation check== | + | ===Aggregation check=== |
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#Prepare 2~5 ml LB liquid medium with appropriate antibiotics into round bottom glass tubes that capped with sponge like cap. Plastic (e.g. polypropyrane) tube is not suitable to earn large cluster of cells. And if you used Erlenmeyer flask, prepare 50 ml LB and antibiotics. | #Prepare 2~5 ml LB liquid medium with appropriate antibiotics into round bottom glass tubes that capped with sponge like cap. Plastic (e.g. polypropyrane) tube is not suitable to earn large cluster of cells. And if you used Erlenmeyer flask, prepare 50 ml LB and antibiotics. | ||
#Add L-arabinose. Final concentration of L-arabinose becomes 1%. | #Add L-arabinose. Final concentration of L-arabinose becomes 1%. | ||
#Suspend colonies into the medium. | #Suspend colonies into the medium. | ||
#Incubate at 34~37C for 24 hrs at 180 rpm. If you wanted to earn large cluster in Erlenmeyer flask, set the temperature at 34C | #Incubate at 34~37C for 24 hrs at 180 rpm. If you wanted to earn large cluster in Erlenmeyer flask, set the temperature at 34C | ||
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==Aggregation check and eCFP expression in low copy plasmid== | ==Aggregation check and eCFP expression in low copy plasmid== | ||
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#Prepare 5 ml LB liquid medium with appropriate antibiotics into round bottom glass tubes that capped with sponge like cap. | #Prepare 5 ml LB liquid medium with appropriate antibiotics into round bottom glass tubes that capped with sponge like cap. | ||
#Suspended colonies into the medium. | #Suspended colonies into the medium. | ||
#Incubated at 37C for 24 hrs at 180 rpm. For this incubation, pipette 150 ul of this cultivating medium to 96-well microtiter plate every 2 hrs. | #Incubated at 37C for 24 hrs at 180 rpm. For this incubation, pipette 150 ul of this cultivating medium to 96-well microtiter plate every 2 hrs. | ||
#Analyzed the Absorbance and fluorescence intensity by microtiter plate reader and fluorescence imager. | #Analyzed the Absorbance and fluorescence intensity by microtiter plate reader and fluorescence imager. | ||
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{{Team:HokkaidoU_Japan/footer}} | {{Team:HokkaidoU_Japan/footer}} | ||
Revision as of 18:34, 26 September 2012
Aggregation Protocols
Aggregation check
- Prepare 2~5 ml LB liquid medium with appropriate antibiotics into round bottom glass tubes that capped with sponge like cap. Plastic (e.g. polypropyrane) tube is not suitable to earn large cluster of cells. And if you used Erlenmeyer flask, prepare 50 ml LB and antibiotics.
- Add L-arabinose. Final concentration of L-arabinose becomes 1%.
- Suspend colonies into the medium.
- Incubate at 34~37C for 24 hrs at 180 rpm. If you wanted to earn large cluster in Erlenmeyer flask, set the temperature at 34C
Aggregation check and eCFP expression in low copy plasmid
- Prepare 5 ml LB liquid medium with appropriate antibiotics into round bottom glass tubes that capped with sponge like cap.
- Suspended colonies into the medium.
- Incubated at 37C for 24 hrs at 180 rpm. For this incubation, pipette 150 ul of this cultivating medium to 96-well microtiter plate every 2 hrs.
- Analyzed the Absorbance and fluorescence intensity by microtiter plate reader and fluorescence imager.
