Team:TU-Delft/Protocol 3
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- | + | <h2 align="center">Making glycerol stocks</h2> | |
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<h4 align="left">Requirements:</h4> | <h4 align="left">Requirements:</h4> | ||
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- | + | <li>bacterial culture </li> | |
- | + | <li>80% glycerol </li> | |
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- | <h4 align="left"> | + | <h4 align="left">Procedure:</h4> |
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- | + | <li> The bacterial cells are grown overnight. </li> | |
+ | <li> In a glass tube take 250 μL of 80 % Glycerol solution. </li> | ||
+ | <li> Pipet 1mL of the cell culture into 0.25 mL 80% glycerol and mix by vortexing and save in -80 °C freezer. </li> | ||
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The glycerol stock can be used whenever required, by just adding 0.5 mL of stock into 5 mL of freshly prepared media. | The glycerol stock can be used whenever required, by just adding 0.5 mL of stock into 5 mL of freshly prepared media. | ||
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Latest revision as of 15:27, 3 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:
- bacterial culture
- 80% glycerol
Procedure:
- The bacterial cells are grown overnight.
- In a glass tube take 250 μL of 80 % Glycerol solution.
- Pipet 1mL of the cell culture into 0.25 mL 80% glycerol and mix by vortexing and save in -80 °C freezer.
The glycerol stock can be used whenever required, by just adding 0.5 mL of stock into 5 mL of freshly prepared media.