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Team:TU-Delft/Protocol2
From 2013.igem.org
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
2. Growing the Single Colonies from the Agar Plates:
Requirements:
1. LB Broth 2. Pipettes 3. Antibiotics 4. Sterile flasksProdecure:
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.
