Team:TU-Delft/Protocol2
From 2013.igem.org
(Difference between revisions)
Line 50: | Line 50: | ||
<li> <a href="https://2013.igem.org/Team:TU-Delft/Protocol_9" style="text-decoration: none""><font | <li> <a href="https://2013.igem.org/Team:TU-Delft/Protocol_9" style="text-decoration: none""><font | ||
color="#0080FF" size="3"> Tricine Gels</font></a> </li> | color="#0080FF" size="3"> Tricine Gels</font></a> </li> | ||
+ | |||
+ | <li> <a href="https://2013.igem.org/Team:TU-Delft/Protocol_10" style="text-decoration: none""><font color="#0080FF" size="3">General Peptide Production</font></a> </li> | ||
</ul> | </ul> |
Revision as of 12:22, 1 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.
- Transforming Parts from Distribution kit
- Growing the Single Colonies from the Agar Plates
- Making glycerol stocks
- Miniprep Protocol
- Restriction digestion
- Ligation
- Gel Extraction Procedure
- PCR Purification
- Tricine Gels
- General Peptide Production
Growing the Single Colonies from the Agar Plates
Requirements:
- LB Broth
- Pipettes
- Antibiotics
- Sterile flasks
Prodecure:
- Prepare flask with LB Broth in it. Add the appropriate antibiotic needed.
- Select the single colony using the pipette tip from the Agar plate, which contains the bacterial cells.
- Inoculate the LB broth with the bacterial cells.
- 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.