IGEM:MIT/2007/Notebook/2007-7-30: Difference between revisions
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(New page: ==Mer-GFP Expression Protocol== # Mix Diluted LB Media (LB4.10) ##4g NaCl, .5g Yeast Exract, 1g Tryptone + 1L H20 # Autoclave LB4.10 # Grow up 10 mls ON at 30°C # Dilute back 1:100 in fr...) |
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Line 4: | Line 4: | ||
# Autoclave LB4.10 | # Autoclave LB4.10 | ||
# Grow up 10 mls ON at 30°C | # Grow up 10 mls ON at 30°C | ||
# Dilute back 1:100 in fresh LB4.10 | # Dilute back 1:100 in fresh LB4.10 (50 ul to 5 ml) | ||
# Add HgCl2 in following concentrations (ng/ml): | # Add HgCl2 in following concentrations (ng/ml): | ||
##0 | ##0 | ||
Line 19: | Line 19: | ||
##10000 | ##10000 | ||
# Grow at 30°C for 16h | # Grow at 30°C for 16h | ||
# Wash a 3ml sample of each (twice) and resuspend in .9% NaCl (Minimizes Background) | |||
# Measure flourescence | |||
* Excitation wavelength - 395nm | |||
* Emission wavelength - 509nm |
Revision as of 09:34, 30 July 2007
Mer-GFP Expression Protocol
- Mix Diluted LB Media (LB4.10)
- 4g NaCl, .5g Yeast Exract, 1g Tryptone + 1L H20
- Autoclave LB4.10
- Grow up 10 mls ON at 30°C
- Dilute back 1:100 in fresh LB4.10 (50 ul to 5 ml)
- Add HgCl2 in following concentrations (ng/ml):
- 0
- 5
- 10
- 25
- 50
- 75
- 100
- 150
- 300
- 500
- 1000
- 10000
- Grow at 30°C for 16h
- Wash a 3ml sample of each (twice) and resuspend in .9% NaCl (Minimizes Background)
- Measure flourescence
- Excitation wavelength - 395nm
- Emission wavelength - 509nm