Splitting 96-well master plate

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When your genomic and expansion plates are confluent, your can split them 1:2 for back ups. 

For the expansion plate have 2 96-well feeder plates prepared ahead of time.  For the genomic preps, have 2 96-well gelatin coated plates ready.  For genomic preps you do not need feeder cells, and this way no feeder DNA will be involved in your Southern.

1. Feed cells 2 hours before splitting, and have new 96-well plates ready with 100 ul of ES media/well
2. Aspirate off media and wash cells with PBS
3. Add 30ul of trypsin (0.05%) and incubate for 5 min. at 37 C
4. Using multichannel pipettor, add 100 ul of ES media and titerate 5-10X
5. Transfer 1/2 the contents (115 ul) to each new 96-well plate.

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