Team:TU-Delft/Protocol 3
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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> |
Revision as of 12:43, 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
- Plasmid Purification Protocol
- Restriction digestion
- Ligation
- Gel Extraction Procedure
- PCR Purification
- Tricine Gels
- General Peptide Production
- SUMO cleavage
- Lysis Protocol
- AIP Sensing Protocol
- Gene Design
- Primer Design
Making glycerol stocks
Requirements:
- Glycerol
- Pipettes
- 1.5 mL Tubes
- Culture Sample
Prodecure:
- Label the tubes and add 180μL of glycerol.
- Pipette out 1000 μL of the culture.
- Add the culture to the tubes and store in -80°C
The glycerol stock can be used whenever required, by just adding 0.5 mL of stock into 5 mL of freshly prepared media.