Picking ES Cell Colonies and
Transferring Them to 96-Well PLATES
1. Add 20 - 25 ul of trypsin per well to a 96-well plate,
round bottom plate.
2. After flooding the 10 cm plate with PBS, pick the colony
from the plate using a Pipetman set at 2 - 4 ul, and transfer it
into the well with trypsin of 96-well plate (1 colony/well).
3. After completing a plate (96 colonies picked), incubate the
plate in TC incubator at 37 �C for 10 - 15 minutes.
4. Add 30 - 35 ul of media, M15 per well. Pipette up and
down to break up the colonies using the multi-channel pipetor.
5. Transfer the ES cell suspension into a 96-well feeder
plate. Add an additional 100 ul of fresh M15 media per well.
Return plates to TC incubator 37� C.
6. Alternatively, 50% of the cells may be lysed directly
for PCR at this point. Add cell suspension directly to PCR Lysis