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Transforming into Bacillus subtillis
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Riachalder (Talk | contribs) (New page: * Add 10µl plasmid to 400µl cells. * Mix by inverting the tube 5 times. * Incubate at 37°C for 60 minutes whilst shaking. * Centrifuge tube at 13,000g for 2 minutes to pellet the cells....) |
Riachalder (Talk | contribs) |
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* Add 10µl plasmid to 400µl cells. | * Add 10µl plasmid to 400µl cells. | ||
* Mix by inverting the tube 5 times. | * Mix by inverting the tube 5 times. | ||
| - | * Incubate at 37°C for 60 minutes whilst shaking. | + | * Incubate at 37°C for 60 minutes whilst shaking. As the tubes are small it is best to leave them shaking on their sides so the liquid mixes properly. To do this, tape the tubes to the rack and place the rack on its side on the shaking table (see diagram). |
| + | [[Image:amazing tube rack.jpg]] | ||
* Centrifuge tube at 13,000g for 2 minutes to pellet the cells. | * Centrifuge tube at 13,000g for 2 minutes to pellet the cells. | ||
* Remove and discard 300µl supernatent. Re-centrifuge if pellet has already resuspended. | * Remove and discard 300µl supernatent. Re-centrifuge if pellet has already resuspended. | ||
* Pipette remaining liquid and pellet up and down to resuspend. | * Pipette remaining liquid and pellet up and down to resuspend. | ||
* Plate onto agar. | * Plate onto agar. | ||
Revision as of 13:10, 10 September 2008
- Add 10µl plasmid to 400µl cells.
- Mix by inverting the tube 5 times.
- Incubate at 37°C for 60 minutes whilst shaking. As the tubes are small it is best to leave them shaking on their sides so the liquid mixes properly. To do this, tape the tubes to the rack and place the rack on its side on the shaking table (see diagram).
- Centrifuge tube at 13,000g for 2 minutes to pellet the cells.
- Remove and discard 300µl supernatent. Re-centrifuge if pellet has already resuspended.
- Pipette remaining liquid and pellet up and down to resuspend.
- Plate onto agar.
