Team:TU-Delft/Protocol 3

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<h2 align="center">Protocols</h2>
 
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Our project deals with <i>E.coli</i> cells which sense Auto-inducing peptides (AIPs) from the <i>Staphylococcus aureus</i> and starts producing Antimicrobial peptides in order to kill the <i>Staphylococcus aureus</i>. Different protocols used during the project are described below.
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  <li> <a href="https://2013.igem.org/Team:TU-Delft/Protocol_1" style="text-decoration: none""><font color="#0080FF" size="3">Transforming Parts from Distribution kit</font></a> </li>
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<li> <a href="https://2013.igem.org/Team:TU-Delft/Protocol2" style="text-decoration: none""><font color="#0080FF" size="3">Growing the Single Colonies from the Agar Plates</font></a> </li>
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<li> <a href="https://2013.igem.org/Team:TU-Delft/Protocol_3" style="text-decoration: none""><font
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color="#0080FF" size="3"> Making glycerol stocks</font></a> </li>
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<li> <a href="https://2013.igem.org/Team:TU-Delft/Protocol_4" style="text-decoration: none""><font
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color="#0080FF" size="3"> Miniprep Protocol</font></a> </li>
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<li> <a href="https://2013.igem.org/Team:TU-Delft/Protocol_5" style="text-decoration: none""><font
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color="#0080FF" size="3"> Restriction digestion</font></a> </li>
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<li> <a href="https://2013.igem.org/Team:TU-Delft/Protocol_6" style="text-decoration: none""><font
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color="#0080FF" size="3"> Ligation</font></a> </li>
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<li> <a href="https://2013.igem.org/Team:TU-Delft/Protocol_7" style="text-decoration: none"" ><font
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color="#0080FF" size="3"> Gel Extraction Procedure</font></a> </li>
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<li> <a href="https://2013.igem.org/Team:TU-Delft/Protocol_8" style="text-decoration: none""><font
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color="#0080FF" size="3">  PCR Purification</font></a> </li>
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<li> <a href="https://2013.igem.org/Team:TU-Delft/Protocol_9" style="text-decoration: none""><font
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color="#0080FF" size="3">  Tricine Gels</font></a> </li>
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<h2 align="center">Making glycerol stocks</h2>
<h2 align="center">Making glycerol stocks</h2>
<h4 align="left">Requirements:</h4>
<h4 align="left">Requirements:</h4>
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<li> Glycerol</li>
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<li>bacterial culture </li>
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<li> Pipettes</li>
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<li>80% glycerol  </li>
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<li> 1.5 mL Tubes</li>
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<li>      Culture Sample</li>
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<h4 align="left">Prodecure:</h4>
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<h4 align="left">Procedure:</h4>
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<li> Label the tubes and add 180μL of glycerol. </li>
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<li> Pipette out 1000 μL of the culture. </li>
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    <li> The bacterial cells are grown overnight. </li>
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<li> Add the culture to the tubes and store in -80°C</li>
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    <li> In a glass tube take 250 μL of 80 % Glycerol solution. </li>
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    <li> Pipet 1mL of the cell culture into 0.25 mL 80% glycerol and mix by vortexing and save in -80 °C freezer.  </li>
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Latest revision as of 15:27, 3 October 2013



Protocols

Our project deals with E.coli cells which sense Auto-inducing peptides (AIPs) from the Staphylococcus aureus and starts producing Antimicrobial peptides in order to kill the Staphylococcus aureus. Different protocols used during the project are described below.




Making glycerol stocks

Requirements:

  1. bacterial culture
  2. 80% glycerol

Procedure:

  1. The bacterial cells are grown overnight.
  2. In a glass tube take 250 μL of 80 % Glycerol solution.
  3. Pipet 1mL of the cell culture into 0.25 mL 80% glycerol and mix by vortexing and save in -80 °C freezer.

The glycerol stock can be used whenever required, by just adding 0.5 mL of stock into 5 mL of freshly prepared media.

Retrieved from "http://2013.igem.org/Team:TU-Delft/Protocol_3"