Team:TU-Delft/Protocol2

From 2013.igem.org

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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>
  <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"" target="_blank"><font  
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  <li> <a href="https://2013.igem.org/Team:TU-Delft/Protocol_3" style="text-decoration: none""><font  
color="#0080FF" size="3"> Making glycerol stocks</font></a> </li>
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"" target="_blank"><font  
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  <li> <a href="https://2013.igem.org/Team:TU-Delft/Protocol_4" style="text-decoration: none""><font  
color="#0080FF" size="3"> Miniprep Protocol</font></a> </li>
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"" target="_blank"><font  
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  <li> <a href="https://2013.igem.org/Team:TU-Delft/Protocol_5" style="text-decoration: none""><font  
color="#0080FF" size="3"> Restriction digestion</font></a> </li>
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  
color="#0080FF" size="3"> Ligation</font></a> </li>
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"" target="_blank"><font  
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  <li> <a href="https://2013.igem.org/Team:TU-Delft/Protocol_7" style="text-decoration: none"" ><font  
color="#0080FF" size="3"> Gel Extraction Procedure</font></a> </li>
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"" target="_blank"><font  
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  <li> <a href="https://2013.igem.org/Team:TU-Delft/Protocol_8" style="text-decoration: none""><font  
color="#0080FF" size="3">  PCR Purification</font></a> </li>
color="#0080FF" size="3">  PCR Purification</font></a> </li>

Revision as of 13:40, 27 September 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.


Growing the Single Colonies from the Agar Plates

Requirements:

1. LB Broth 2. Pipettes 3. Antibiotics 4. Sterile flasks

Prodecure:

1. Prepare flask with LB Broth in it. Add the appropriate antibiotic needed.
2. Select the single colony using the pipette tip from the Agar plate, which contains the bacterial cells.
3. Inoculate the LB broth with the bacterial cells.
3. Grow the culture for 14-16 hours.

Next day prepare glycerol stocks and carry out a miniprep protocol to extract the desired plasmid from the bacterial cells.