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

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(Mer-GFP Expression Protocol)
Line 23: Line 23:
* Excitation wavelength - 395nm
* Excitation wavelength - 395nm
* Emission wavelength - 509nm
* Emission wavelength - 509nm
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 +
==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

Revision as of 14:22, 30 July 2007

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

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