Separation of oligonucleosomal DNA fragments from apoptotic animal cells using a triad of Sephacryl columns.

The degradation of DNA, resulting in information of oligonucleosomal fragments, is a characteristic feature of apoptosis, i.e., the process of programmed cell death. In this work we have developed a method for exact determination of the proportion of fragmented DNA in an apoptotic cell population. To this end we employed Sephacryl gel chromatography matrices and UV detection of DNA concentration. The disturbing effect of low-molecular-mass UV-absorbing contaminants was eliminated by insertion of a Sephacryl S-200 HR pre-column. Optimum resolution of DNA samples isolated from apoptotic cells was achieved using a triad of Sephacryl S-200 HR, Sephacryl S-500 HR and Sephacryl S-1000 SF columns.