Transforming into Bacillus subtillis

Revision as of 10:51, 18 September 2008 by Riachalder (Talk | contribs)

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.