Difference between revisions of "Team:Grenoble-Alpes/Protocols"
| Line 172: | Line 172: | ||
Materials | Materials | ||
<li>20mL LB Agar</li> | <li>20mL LB Agar</li> | ||
| − | <li> | + | <li>20µL Cam (25µg/mL)</li> |
<br> | <br> | ||
Methods | Methods | ||
| Line 243: | Line 243: | ||
Methods | Methods | ||
<ol> | <ol> | ||
| − | <li>Dissolve 0,171g of sucrose in 5mL of sterile water<li> | + | <li>Dissolve 0,171g of sucrose in 5mL of sterile water</li> |
<li>Conserve at 4°C</li> | <li>Conserve at 4°C</li> | ||
</ol> | </ol> | ||
| Line 254: | Line 254: | ||
<h5> | <h5> | ||
Materials | Materials | ||
| − | <li> | + | <li>80µL IPTG (500mM)</li> |
<li>10mL LB</li> | <li>10mL LB</li> | ||
<li>10µL Cam (25ug/mL)</li> | <li>10µL Cam (25ug/mL)</li> | ||
| Line 260: | Line 260: | ||
Methods | Methods | ||
<ol> | <ol> | ||
| − | <li>Add | + | <li>Add 10µL of Cam in 10mL of LB</li> |
<li>Homogenize</li> | <li>Homogenize</li> | ||
| − | <li>Remove | + | <li>Remove 90µL of solution and add 80µL of IPTG</li> |
<li>Conserve at -20°C</li> | <li>Conserve at -20°C</li> | ||
</ol> | </ol> | ||
Revision as of 07:25, 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 10µL of Cam in 10mL of LB
- Homogenize
- Remove 90µL of solution and add 80µL of IPTG
- Conserve at -20°C
CLOSE
Experimentations
