Team:UEA-JIC Norwich/algaeelectroporation

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University of East Anglia-JIC

UNIVERSITY OF EAST ANGLIA-JOHN INNES CENTRE

1. Transfer 50ml of algae culture into four 50ml screw top tubes. <p style="color:#000000">2. Place all four tubes in the eppendorf centrifuge at 800rpm for 9 minutes at 20-22°C. <p style="color:#000000">3. Immediately pour contents of tubes into glass beaker and keep tubes upside down so no unwanted suspension mixes with the pellet. <p style="color:#000000">4. For all tubes resuspend cells in 250µl of TAP and sucrose media. <p style="color:#000000">5. Add linearised G-Luc plasmid (5µL)to the suspension and place on ice. <p style="color:#000000">6. Transfer 250µL of algae suspension into 1mL cuvette. <p style="color:#000000">7. Subject cuvettes to electroporation at 2200v/ cm and place back on ice. <p style="color:#000000">8. Add 500µL of starch solution to each cuvette and aspirate gently. <p style="color:#000000">9. Add 750µL of each sample onto an LB plate. <p style="color:#000000">10.Parafilm the lid and plate to seal and place in a light incubator.

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