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Team:Calgary/Notebook/Protocols/Process6
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Notebook
Post-Regionals
Journal
- Defining Objectives
- Techniques Workshop
- Bioinformatics & qRT-PCR
- Sensory Element Fishing
- Electrochem - CPR Oxidation
- Electrochem - Reporter Gene
- Pseudomonas Tools
- Microalgae Tools
- NA Viability Assays
- May - July Group Journal
Protocols
Safety
Plasmid Extraction
Reagents and Materials
Protocol
- A solution containing 20% (w/v) PEG was prepared and then added to cells of D. salina before transformation.
- Glass beads, 0.45– 0.52 mm in diameter, were washed with concentrated sulfuric acid, then rinsed thoroughly with distilled water, and dried by baking at 180°C for 2–3 h.
- D. salina cells were cultured to logarithmic phase and then harvested by centrifugation at 5,000 rpm for 2 min.
- Cells were washed three times with the liquid medium as mentioned above, and resuspended with this medium at a concentration of 105 cells/ml.
- A mixture with 300 mg glass beads, 40 µg vector DNA (0.4 µg/µl) and 100 µl PEG was added to 0.8 ml of D. salina cell culture, mixed briefly by gently inverting tube and then agitated at 2,000 rpm on a vortex for 6 sec in 1.5 ml centrifuge tubes.
- The glass beads were allowed to settle, and cells were transferred to sterilized test tubes and cultured in liquid medium under dim light condition for 24 h.
