Difference between revisions of "Team:Bielefeld-CeBiTec/Media"
| Line 133: | Line 133: | ||
<div class="panel-heading"> | <div class="panel-heading"> | ||
<h4 class="panel-title"> | <h4 class="panel-title"> | ||
| − | <a data-toggle="collapse" href="#collapseFive"> | + | <a data-toggle="collapse" href="#collapseFive">Equilibration buffer for lacI purification</a> |
</h4> | </h4> | ||
</div> | </div> | ||
| Line 141: | Line 141: | ||
<li>For each litre of solution: </li> | <li>For each litre of solution: </li> | ||
<ul> | <ul> | ||
| − | <li> | + | <li>29.22g NaCl </li> |
| − | <li> | + | <li>1.36 g imidazole </li> |
| − | <li> | + | <li>29 ml of 86 % glycerol </li> |
</ul> <br> | </ul> <br> | ||
| − | <li> | + | <li>Add all these ingredients to a 50 mM sodium phosphate buffer (pH 8.0). Prior to use add DTT to a final concentration of 1mM. </li> |
| − | + | ||
| − | + | ||
<li>store at room temperature. </li> <br> | <li>store at room temperature. </li> <br> | ||
<li>Note: Final (1x) working concentration: </li> | <li>Note: Final (1x) working concentration: </li> | ||
<ul> | <ul> | ||
| − | <li> | + | <li>50 mM sodium phosphate </li> |
| − | <li> | + | <li>500 mM NaCl </li> |
| + | <li>20 mM imidazole</li> | ||
| + | <li>2.5 % glycerol </li> | ||
| + | <li>1 mM DTT</li> | ||
</ul> | </ul> | ||
</ul> | </ul> | ||
| Line 161: | Line 163: | ||
<div class="panel-heading"> | <div class="panel-heading"> | ||
<h4 class="panel-title"> | <h4 class="panel-title"> | ||
| − | <a data-toggle="collapse" href="#collapseSix"> | + | <a data-toggle="collapse" href="#collapseSix">Elution buffer for purification of lacI</a> |
</h4> | </h4> | ||
</div> | </div> | ||
| Line 169: | Line 171: | ||
<li>For each litre of solution: </li> | <li>For each litre of solution: </li> | ||
<ul> | <ul> | ||
| − | <li> | + | <li>17.53 g NaCl </li> |
| − | <li> | + | <li>13.6 g imidazole </li> |
| − | + | ||
</ul> <br> | </ul> <br> | ||
| − | + | <li>Add all these ingredients to a 20 mM sodium phosphate buffer (pH 7.4). </li> | |
| − | + | ||
| − | + | ||
<li>store at room temperature. </li> <br> | <li>store at room temperature. </li> <br> | ||
<li>Note: Final (1x) working concentration: </li> | <li>Note: Final (1x) working concentration: </li> | ||
<ul> | <ul> | ||
| − | <li> | + | <li>20 mM sodium phosphate</li> |
| − | <li> | + | <li>300 mM NaCl </li> |
| + | <li>200 mM imidazole</li> | ||
| + | </ul> | ||
| + | </ul> | ||
| + | </div> | ||
| + | </div> | ||
| + | </div> | ||
| + | <div class="panel panel-default"> | ||
| + | <div class="panel-heading"> | ||
| + | <h4 class="panel-title"> | ||
| + | <a data-toggle="collapse" href="#collapseSeven">Buffer for dialysis of lacI</a> | ||
| + | </h4> | ||
| + | </div> | ||
| + | <div id="collapseSeven" class="panel-collapse collapse"> | ||
| + | <div class="panel-body"> | ||
| + | <ul> | ||
| + | <li>For each litre of solution: </li> | ||
| + | <ul> | ||
| + | <li>50 g glucose </li> | ||
| + | |||
| + | </ul> <br> | ||
| + | <li>Add the glucose to a 200 mM potassium phosphate buffer (pH 7.6). Prior to usage add DTT to a final concentration of 1 mM</li> | ||
| + | <li>store at room temperature. </li> <br> | ||
| + | <li>Note: Final (1x) working concentration: </li> | ||
| + | <ul> | ||
| + | <li>200 mM potassium phosphate</li> | ||
| + | <li>5 % glucose </li> | ||
| + | <li>1 mM DTT</li> | ||
</ul> | </ul> | ||
</ul> | </ul> | ||
Revision as of 20:17, 6 July 2015
Media
- For 1 liter LB:
- 20 g LB powder
- Fill the bottle with deionized H2O
- For 1 liter LB-plates:
- 18 g LB powder
- 16 g Select Agar
- Fill the bottle with deionized H2O
- Add the following components for 900 ml of distilled H2O:
- 20 g Trypton
- 5 g Bacto Yeast Extract
- 2 ml of 5 M NaCl
- 2.5 ml of 1 M KCl
- 10 ml of 1 M MgCl2
- 10 ml of 1 M MgSO4
- 20 ml of 1 M glucose
Buffers
- For each litre of solution:
- 242 g Tris Base (MW=121.1)
- 57.1 ml Glacial Acetic Acid
- 100 ml 0.5 M EDTA
- mix Tris with stir bar to dissolve in about 600 ml of ddH2O.
- add the EDTA and Acetic Acid.
- bring final volume to 1 l with ddH20.
- store at room temperature.
- Note: Final (1x) working concentration:
- 0.04 M Tris - Acetate
- 0.001 M EDTA
- For each 200 ml of solution:
- 5.844 g NaCl
- 0.272 g imidazole
- 5.8 ml of 86 % glycerol
- 0.406 g MgCl2
- 0.2 g lysozyme
- Add all these ingredients to a 50 mM sodium phosphate buffer (pH 8.0). Prior to use add DTT to a final concentration of 1mM and tween 20 to a final concentration of 0.1 %.
- store at room temperature.
- Note: Final (1x) working concentration:
- 50 mM Sodiumphosphate
- 500 mM NaCl
- 20 mM imidazole
- 2.5 % glxcerol
- 1 mM DTT
- 10 mM MgCl2
- 0.1 % tween 20
- 1 mg/ml lysozyme
- For each litre of solution:
- 29.22g NaCl
- 1.36 g imidazole
- 29 ml of 86 % glycerol
- Add all these ingredients to a 50 mM sodium phosphate buffer (pH 8.0). Prior to use add DTT to a final concentration of 1mM.
- store at room temperature.
- Note: Final (1x) working concentration:
- 50 mM sodium phosphate
- 500 mM NaCl
- 20 mM imidazole
- 2.5 % glycerol
- 1 mM DTT
- For each litre of solution:
- 17.53 g NaCl
- 13.6 g imidazole
- Add all these ingredients to a 20 mM sodium phosphate buffer (pH 7.4).
- store at room temperature.
- Note: Final (1x) working concentration:
- 20 mM sodium phosphate
- 300 mM NaCl
- 200 mM imidazole
- For each litre of solution:
- 50 g glucose
- Add the glucose to a 200 mM potassium phosphate buffer (pH 7.6). Prior to usage add DTT to a final concentration of 1 mM
- store at room temperature.
- Note: Final (1x) working concentration:
- 200 mM potassium phosphate
- 5 % glucose
- 1 mM DTT