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12. Centrifuge the tubes at 13000g for 30 seconds and remove the supernatant.
A. *For prototrophic gene selection: *Pipette 200 uL - 1.0 ml of sterile water into the transformation tube. Stir the pellet with a sterile micropipette tip to break the cell pellet and then vortex mix to uniformly resuspend pellet. **{}I usually resuspend in 200 uL.*
B. *For eukaryotic antibiotic gene selection: *Pipette 200 uL - 1.0 ml of YPD liquid medium into the transformation tube. Vortex mix to resuspend pellet. Incubate for 2--3 h at 30C to ensure good expression from the input plasmid DNA.
13. Plate 200 uL of the cell suspension on the appropriate plates. Incubate plates at 30C for 2-3 days.Prototrophic gene selection
(i) Pipette 1.0 ml of sterile water into the transformation tube. Stir the pellet with a sterile micropipette tip to break the cell
pellet and then vortex mix to uniformly resuspend pellet.
(B) Eukaryotic antibiotic gene selection
(i) Pipette 1.0 ml of YPD liquid medium into the transformation tube. Vortex mix to resuspend pellet.
(ii) Incubate for 2--3 h at 30 1C to ensure good expression from the input plasmid DNA.