Metaphase chromosomes with high molecular weight DNA were isolated from Chinese hamster ovary (CHO) cells in a neutral buffer containing polyamines and chelators. The individual, unfixed chromosomes retained their centromeric and secondary constrictions, distinct sister chromatids, and complex banding patterns. The DNA from these chromosomes was 100-fold larger (2 x 10(8) daltons) than DNA from chromosomes isolated by other procedures. These characteristics indicate preservation during isolation of considerable native structure. In contrast to chromosomes produced by other methods, these chromosomes were stable in storage and did not aggregate, thus providing useful material for studies of the structure and biochemistry of individual chromosomes.
Article|
April 01 1979
Rapid isolation of metaphase chromosomes containing high molecular weight DNA.
A B Blumenthal
J D Dieden
L N Kapp
J W Sedat
Online ISSN: 1540-8140
Print ISSN: 0021-9525
J Cell Biol (1979) 81 (1): 255–259.
Citation
A B Blumenthal, J D Dieden, L N Kapp, J W Sedat; Rapid isolation of metaphase chromosomes containing high molecular weight DNA.. J Cell Biol 1 April 1979; 81 (1): 255–259. doi: https://doi.org/10.1083/jcb.81.1.255
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