IGEM:MIT/2007/Notebook/2007-7-30

Mer-GFP Expression Protocol

1. Mix Diluted LB Media (LB4.10)
   1. 4g NaCl, .5g Yeast Exract, 1g Tryptone + 1L H20
2. Autoclave LB4.10
3. Grow up 10 mls ON at 30°C
4. Dilute back 1:100 in fresh LB4.10 (50 ul to 5 ml)
5. Add HgCl2 in following concentrations (ng/ml):
   1. 0
   2. 5
   3. 10
   4. 25
   5. 50
   6. 75
   7. 100
   8. 150
   9. 300
   10. 500
   11. 1000
   12. 10000
6. Grow at 30°C for 16h
7. Wash a 3ml sample of each (twice) and resuspend in .9% NaCl (Minimizes Background)
8. Measure flourescence

• Excitation wavelength - 395nm
• Emission wavelength - 509nm

==Measure out 0.1 g and dilute in 100 mL water This will give you 100 mL of 1 mg/mL Take 10 uL and dilute in 10 mL water This will give you 10 mL of 1 ug/mL Take 1 mL and dilute in 10 mL water This will give you 10 mL of 100 ng/mL