Difference between revisions of "Team:Grenoble-Alpes/Protocols"
| Line 124: | Line 124: | ||
<div id="lb_broth"> | <div id="lb_broth"> | ||
<h5> | <h5> | ||
| − | Materials | + | Materials |
<li>20g LB Broth Base (powder)</li> | <li>20g LB Broth Base (powder)</li> | ||
<li>1L Distillated Water</li> | <li>1L Distillated Water</li> | ||
| − | Methods | + | Methods |
<ol> | <ol> | ||
<li>Add 20g of LB Broth Base per 1L of distillated water</li> | <li>Add 20g of LB Broth Base per 1L of distillated water</li> | ||
| Line 139: | Line 139: | ||
<div id="lb_agar"> | <div id="lb_agar"> | ||
<h5> | <h5> | ||
| − | Materials | + | Materials |
<li>32g LB Agar (powder)</li> | <li>32g LB Agar (powder)</li> | ||
<li>1L Distilled Water</li> | <li>1L Distilled Water</li> | ||
| − | Methods | + | Methods |
<ol> | <ol> | ||
<li>Add 32g of LB Agar per 1L of distilled water</li> | <li>Add 32g of LB Agar per 1L of distilled water</li> | ||
| Line 154: | Line 154: | ||
<div id="chloramphenicol"> | <div id="chloramphenicol"> | ||
<h5> | <h5> | ||
| − | + | Materials | |
| + | <li>0,25g Chloramphenicol (25mg/mL)</li> | ||
| + | <li>10mL Ethanol</li> | ||
| + | Methods | ||
| + | <ol> | ||
| + | <li>Add 0,25g of Cam per 10mL of ethanol</li> | ||
| + | <li>Homogenize by vortexing</li> | ||
| + | <li>Filter with a 22mm membrane filter</li> | ||
| + | <li>Conserve at -20°C in 1mL aliquot</li> | ||
| + | </ol> | ||
</h5> | </h5> | ||
<a href="#prep" style="text-decoration: none;"><h2 class="top">TOP</h2></a> | <a href="#prep" style="text-decoration: none;"><h2 class="top">TOP</h2></a> | ||
| Line 161: | Line 170: | ||
<div id="petri_dish"> | <div id="petri_dish"> | ||
<h5> | <h5> | ||
| − | + | Materials | |
| + | <li>20mL LB Agar</li> | ||
| + | <li>20uL Cam (25ug/mL)</li> | ||
| + | Methods | ||
| + | <ol> | ||
| + | <li>Add of 20uL Cam in 20mL of liquid LB Agar</li> | ||
| + | <li>Pour the solution in the plate</li> | ||
| + | <li>Wait until the agar solidifies</li> | ||
| + | <li>Conserve returned in 4°C</li> | ||
| + | </ol> | ||
</h5> | </h5> | ||
<a href="#prep" style="text-decoration: none;"><h2 class="top">TOP</h2></a> | <a href="#prep" style="text-decoration: none;"><h2 class="top">TOP</h2></a> | ||
| Line 168: | Line 186: | ||
<div id="glycerol"> | <div id="glycerol"> | ||
<h5> | <h5> | ||
| − | + | Materials | |
| + | <li>80mL Glycerol 100%</li> | ||
| + | <li>20mL Sterile Water</li> | ||
| + | Methods | ||
| + | <ol> | ||
| + | <li>Harvest 80mL of glycerol 100%</li> | ||
| + | <li>Add 20mL of sterile water</li> | ||
| + | <li>Conserve at 4°C</li> | ||
| + | </ol> | ||
</h5> | </h5> | ||
<a href="#prep" style="text-decoration: none;"><h2 class="top">TOP</h2></a> | <a href="#prep" style="text-decoration: none;"><h2 class="top">TOP</h2></a> | ||
| Line 175: | Line 201: | ||
<div id="cacl2"> | <div id="cacl2"> | ||
<h5> | <h5> | ||
| − | + | Materials | |
| + | <li>0,0555g CaCl2 (111g/mol)</li> | ||
| + | <li>5mL Sterile Water</li> | ||
| + | Methods | ||
| + | <ol> | ||
| + | <li>Dissolve 0,056g of CaCl2 in 5mL of sterile water</li> | ||
| + | <li>Conserve at 4°C</li> | ||
| + | </ol> | ||
</h5> | </h5> | ||
<a href="#prep" style="text-decoration: none;"><h2 class="top">TOP</h2></a> | <a href="#prep" style="text-decoration: none;"><h2 class="top">TOP</h2></a> | ||
| Line 182: | Line 215: | ||
<div id="mgcl2"> | <div id="mgcl2"> | ||
<h5> | <h5> | ||
| − | + | Materials | |
| + | <li>0,610g MgCl2 (203,31g/mol)</li> | ||
| + | <li>30mL Sterile Water</li> | ||
| + | Methods | ||
| + | <ol> | ||
| + | <li>Dissolve 0,6105g of MgCl2 in 30mL of sterile water</li> | ||
| + | <li>Conserve at 4°C</li> | ||
| + | </ol> | ||
</h5> | </h5> | ||
<a href="#prep" style="text-decoration: none;"><h2 class="top">TOP</h2></a> | <a href="#prep" style="text-decoration: none;"><h2 class="top">TOP</h2></a> | ||
| Line 189: | Line 229: | ||
<div id="sucrose"> | <div id="sucrose"> | ||
<h5> | <h5> | ||
| − | + | Materials | |
| + | <li>0,171g Sucrose</li> | ||
| + | <li>5mL Sterile Water</li> | ||
| + | Methods | ||
| + | <ol> | ||
| + | <li>Dissolve 0,171g of sucrose in 5mL of sterile water<li> | ||
| + | <li>Conserve at 4°C</li> | ||
| + | </ol> | ||
</h5> | </h5> | ||
<a href="#prep" style="text-decoration: none;"><h2 class="top">TOP</h2></a> | <a href="#prep" style="text-decoration: none;"><h2 class="top">TOP</h2></a> | ||
| Line 196: | Line 243: | ||
<div id="iptg"> | <div id="iptg"> | ||
<h5> | <h5> | ||
| − | + | Materials | |
| + | <li>80uL IPTG (500mM)</li> | ||
| + | <li>10mL LB</li> | ||
| + | <li>10µL Cam (25ug/mL)</li> | ||
| + | Methods | ||
| + | <ol> | ||
| + | <li>Add 10uL of Cam in 10mL of LB</li> | ||
| + | <li>Homogenize</li> | ||
| + | <li>Remove 90uL of solution and add 80uL of IPTG</li> | ||
| + | <li>Conserve at -20°C</li> | ||
| + | </ol> | ||
</h5> | </h5> | ||
<a href="#prep" style="text-decoration: none;"><h2 class="top">TOP</h2></a> | <a href="#prep" style="text-decoration: none;"><h2 class="top">TOP</h2></a> | ||
Revision as of 15:21, 29 August 2017
Protocols
Preparations
Experimentations
Materials
- Add 20g of LB Broth Base per 1L of distillated water
- Homogenize
- Autoclave at 121°C for 15 minutes
TOP
Materials
- Add 32g of LB Agar per 1L of distilled water
- Homogenize
- Autoclave at 121°C for 15 minutes
TOP
Materials
- 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
TOP
Materials
- 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
TOP
Materials
- Harvest 80mL of glycerol 100%
- Add 20mL of sterile water
- Conserve at 4°C
TOP
Materials
- Dissolve 0,056g of CaCl2 in 5mL of sterile water
- Conserve at 4°C
TOP
Materials
- Dissolve 0,6105g of MgCl2 in 30mL of sterile water
- Conserve at 4°C
TOP
Materials
- Dissolve 0,171g of sucrose in 5mL of sterile water
- Conserve at 4°C
TOP
Materials
- Add 10uL of Cam in 10mL of LB
- Homogenize
- Remove 90uL of solution and add 80uL of IPTG
- Conserve at -20°C
TOP
