metaphase chromosomes

dcoleman at dcoleman at
Wed Mar 10 03:16:47 EST 1993

 dear bionetters,
 I am a graduate student working on DNA sequencing, but as a side project I am 
 interested in subjecting metaphase chromosomes to electron microscopy. I 
 understanding how to arrest the cells in metaphase --I am working with
 a borrowed culture of a human lymphocytes-- but am unclear on how to extract 
 and purify the chromosomes.
 Basically I am looking for two procedures: 
 1. how to break open the cells gently, and 
 2. how to isolate the chromosomes from the remains of the cell. (A few papers 
 mentioned centrifugation, but fewer still provided the exact conditions!)
 Any advice on procedures or even references would be greatly apreciated.

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