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Team:WashU/Protocols/Transformation6803
From 2012.igem.org
(Difference between revisions)
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1. Make sure the cells are in exponential growth phase. (OD730 of 0.8) | 1. Make sure the cells are in exponential growth phase. (OD730 of 0.8) | ||
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2. Take 15 ml of cell culture and spin it down for 15 minutes at RT. | 2. Take 15 ml of cell culture and spin it down for 15 minutes at RT. | ||
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3. ReSuspend the pellet to 1ml in BG11 broth. | 3. ReSuspend the pellet to 1ml in BG11 broth. | ||
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4. Mix in the plasmid to a final concentration of 150 ng/ml | 4. Mix in the plasmid to a final concentration of 150 ng/ml | ||
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5. Incubate for 5 h at 25 degrees C and in the light. | 5. Incubate for 5 h at 25 degrees C and in the light. | ||
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6. Plate the cells onto BG11 plates with antibiotic. | 6. Plate the cells onto BG11 plates with antibiotic. | ||
Spray your gloves, containers, and working surfaces with ethanol, and let the ethanol dry, before | Spray your gloves, containers, and working surfaces with ethanol, and let the ethanol dry, before | ||
handling cyanobacteria. | handling cyanobacteria. | ||
Revision as of 15:54, 14 June 2012
1. Make sure the cells are in exponential growth phase. (OD730 of 0.8)
2. Take 15 ml of cell culture and spin it down for 15 minutes at RT.
3. ReSuspend the pellet to 1ml in BG11 broth.
4. Mix in the plasmid to a final concentration of 150 ng/ml
5. Incubate for 5 h at 25 degrees C and in the light.
6. Plate the cells onto BG11 plates with antibiotic.
Spray your gloves, containers, and working surfaces with ethanol, and let the ethanol dry, before handling cyanobacteria.
