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Protocols: ES Cell Culture - Gene Targeting - Protocols

Picking ES Cell Colonies and
Transferring Them to 24-Well Culture Dishes

	1.	Spot 15 - 25 ul of trypsin in a 24-well array onto the lid 
of a 10 cm plate.

	2.	After flooding the plate with PBS, pick the colony from 
the plate using a Pipetman set at 2 ul, and transfer it into the trypsin.  
Leave in trypsin for 10-15 minutes.

	3.	Add 20 ul of media from the refed feeder plate to the 
trypsin spot.  Pipet up and down to break up the colony.

	4.	Transfer the ES cell suspension (approximately 37 ul) 
into the well of a 24-well feeder plate.

	5.	Alternatively, 50% of the cells may be lysed directly 
for PCR at this point.  Add 15 ul of the cell suspension directly 
to 25 ul of PCR Lysis Buffer.