Difference between revisions of "Team:Bielefeld-CeBiTec/Media"

Line 112: Line 112:
 
                                 <li>0.2 g lysozyme</li>
 
                                 <li>0.2 g lysozyme</li>
 
                             </ul> <br>
 
                             </ul> <br>
−
                                 <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>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 %. </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>0.04 M Tris - Acetate </li>
+
                                         <li>50 mM Sodiumphosphate </li>
−
                                         <li>0.001 M EDTA </li>  
+
                                        <li>500 mM NaCl </li>
 +
<li>20 mM imidazole </li>
 +
                                        <li>2.5 % glxcerol </li>
 +
<li>1 mM DTT </li>
 +
                                         <li>10 mM MgCl<sub>2</sub> </li>
 +
<li>0.1 % tween 20 </li>
 +
                                        <li>1 mg/ml lysozyme</li>
 
                                     </ul>  
 
                                     </ul>  
 
                     </ul>
 
                     </ul>

Revision as of 20:03, 6 July 2015

iGEM Bielefeld 2015


Media

LB

  • 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

SOC

  • 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

50X TAE Stock Solution

  • 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

Lysis and binding buffer for purification of lacI

  • 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

50X TAE Stock Solution

  • 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

50X TAE Stock Solution

  • 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