Team:UEA-JIC Norwich/algaeelectroporation

From 2011.igem.org

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

Latest revision as of 14:53, 21 September 2011

University of East Anglia-JIC

UNIVERSITY OF EAST ANGLIA-JOHN INNES CENTRE

1. Transfer 50ml of algae culture into four 50ml screw top tubes.

2. Place all four tubes in the eppendorf centrifuge at 800rpm for 9 minutes at 20-22°C.

3. Immediately pour contents of tubes into glass beaker and keep tubes upside down so no unwanted suspension mixes with the pellet.

4. For all tubes resuspend cells in 250µl of TAP and sucrose media.

5. Add linearised G-Luc plasmid (5µL)to the suspension and place on ice.

6. Transfer 250µL of algae suspension into 1mL cuvette.

7. Subject cuvettes to electroporation at 2200v/ cm and place back on ice.

8. Add 500µL of starch solution to each cuvette and aspirate gently.

9. Add 750µL of each sample onto an LB plate.

10.Parafilm the lid and plate to seal and place in a light incubator.

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