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Team:TU-Delft/Protocol2
From 2013.igem.org
(Difference between revisions)
| Line 52: | Line 52: | ||
<h2 align="center">Growing the Single Colonies from the Agar Plates</h2> | <h2 align="center">Growing the Single Colonies from the Agar Plates</h2> | ||
<h4 align="left">Requirements:</h4> | <h4 align="left">Requirements:</h4> | ||
| - | + | <ol> | |
| - | + | <li> LB Broth</li> | |
| - | + | <li> Pipettes</li> | |
| - | + | <li> Antibiotics</li> | |
| + | <li> Sterile flasks</li> | ||
| + | </ol> | ||
<br> | <br> | ||
<h4 align="left">Prodecure:</h4> | <h4 align="left">Prodecure:</h4> | ||
| - | + | <ol> | |
| - | + | <li> Prepare flask with LB Broth in it. Add the appropriate antibiotic needed.</li> | |
| - | + | <li> Select the single colony using the pipette tip from the Agar plate, which contains the bacterial cells. </li> | |
| - | + | <li> Inoculate the LB broth with the bacterial cells. </li> | |
| + | <li> Grow the culture for 14-16 hours. </li> | ||
<br> | <br> | ||
Next day prepare glycerol stocks and carry out a miniprep protocol to extract the desired plasmid from the bacterial cells. | Next day prepare glycerol stocks and carry out a miniprep protocol to extract the desired plasmid from the bacterial cells. | ||
Revision as of 12:19, 29 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.
- 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
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.
