ISOLATION OF DNA FROM SEA URCHIN SPERM NUCLEI BY HYDROXYAPATITE CHROMATOGRAPHY

This article describes in detail the procedures in obtaining spermatids from living animals and in isolating sperm nuclei DNA by hydroxyapatite chromatography.The isolated DNA is free of RNAs and proteins after treatment with proteinase K, followed by chromatography on hydroxyapatite column which is eluted with stepwise-increased concentrations of potassium phosphate buffer.More than 89% of the DNA purified is found in the eluates of both 0.2 M and 0.25 M potassium phosphate buffer.The two fractions are identical in ultraviolet absorption spectrum.DNA molecules thus prepared should be useful for studying DNA structure and DNA replication by electron microscopy, since the length of the purified DNA molecules is in the range of the size of a replicating unit.