Difference between revisions of "Team:Grenoble-Alpes/Protocols"
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<li>20g LB Broth Base (powder)</li> | <li>20g LB Broth Base (powder)</li> | ||
<li>1L Distillated Water</li> | <li>1L Distillated Water</li> | ||
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Methods | Methods | ||
<ol> | <ol> | ||
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<li>32g LB Agar (powder)</li> | <li>32g LB Agar (powder)</li> | ||
<li>1L Distilled Water</li> | <li>1L Distilled Water</li> | ||
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Methods | Methods | ||
<ol> | <ol> | ||
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<li>0,25g Chloramphenicol (25mg/mL)</li> | <li>0,25g Chloramphenicol (25mg/mL)</li> | ||
<li>10mL Ethanol</li> | <li>10mL Ethanol</li> | ||
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Methods | Methods | ||
<ol> | <ol> | ||
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<li>20mL LB Agar</li> | <li>20mL LB Agar</li> | ||
<li>20uL Cam (25ug/mL)</li> | <li>20uL Cam (25ug/mL)</li> | ||
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Methods | Methods | ||
<ol> | <ol> | ||
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<li>80mL Glycerol 100%</li> | <li>80mL Glycerol 100%</li> | ||
<li>20mL Sterile Water</li> | <li>20mL Sterile Water</li> | ||
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Methods | Methods | ||
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<li>0,0555g CaCl2 (111g/mol)</li> | <li>0,0555g CaCl2 (111g/mol)</li> | ||
<li>5mL Sterile Water</li> | <li>5mL Sterile Water</li> | ||
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Methods | Methods | ||
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<li>0,610g MgCl2 (203,31g/mol)</li> | <li>0,610g MgCl2 (203,31g/mol)</li> | ||
<li>30mL Sterile Water</li> | <li>30mL Sterile Water</li> | ||
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Methods | Methods | ||
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<li>0,171g Sucrose</li> | <li>0,171g Sucrose</li> | ||
<li>5mL Sterile Water</li> | <li>5mL Sterile Water</li> | ||
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Methods | Methods | ||
<ol> | <ol> | ||
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<li>10mL LB</li> | <li>10mL LB</li> | ||
<li>10µL Cam (25ug/mL)</li> | <li>10µL Cam (25ug/mL)</li> | ||
| + | <br> | ||
Methods | Methods | ||
<ol> | <ol> | ||
Revision as of 07:12, 30 August 2017
Protocols
Preparations
LB Broth
Materials
Materials
Methods
- Add 20g of LB Broth Base per 1L of distillated water
- Homogenize
- Autoclave at 121°C for 15 minutes
CLOSE
LB Agar
Materials
Materials
Methods
- Add 32g of LB Agar per 1L of distilled water
- Homogenize
- Autoclave at 121°C for 15 minutes
CLOSE
Chloramphenicol (Cam) 25ug/mL Stock
Materials
Materials
Methods
- Add 0,25g of Cam per 10mL of ethanol
- Homogenize by vortexing
- Filter with a 22mm membrane filter
- Conserve at -20°C in 1mL aliquot
CLOSE
Petri Dish Preparation
Materials
Materials
Methods
- Add of 20uL Cam in 20mL of liquid LB Agar
- Pour the solution in the plate
- Wait until the agar solidifies
- Conserve returned in 4°C
CLOSE
Glycerol 80%
Materials
Materials
Methods
- Harvest 80mL of glycerol 100%
- Add 20mL of sterile water
- Conserve at 4°C
CLOSE
CaCl2 100mM
Materials
Materials
Methods
- Dissolve 0,056g of CaCl2 in 5mL of sterile water
- Conserve at 4°C
CLOSE
MgCl2 100mM
Materials
Materials
Methods
- Dissolve 0,6105g of MgCl2 in 30mL of sterile water
- Conserve at 4°C
CLOSE
Sucrose 100mM
Materials
Materials
Methods
- Dissolve 0,171g of sucrose in 5mL of sterile water
- Conserve at 4°C
CLOSE
IPTG 4mM
Materials
Materials
Methods
- Add 10uL of Cam in 10mL of LB
- Homogenize
- Remove 90uL of solution and add 80uL of IPTG
- Conserve at -20°C
CLOSE
Experimentations
