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 

Buffer.