Interaction of DNA with hydroxyapatite. Studies on the effect of the phosphate concentration of the column equilibration and washing buffer.
The ability of hydroxyapatite to bind DNA effectively in phosphate solutions used for column equilibration, sample loading and column washing has been examined. It was demonstrated that substantial amounts of DNA (up to 40%) were eluted in the washing buffer when the phosphate concentration in the lysing solution or urea-phosphate used for column equilibration, sample loading and column washing was 0.24 M. A reduction in the phosphate concentration from 0.24 to 0.15 M in urea-phosphate solution led to almost 100% binding, whereas a similar reduction in the lysing solution did not. A modified method for loading and eluting DNA from hydroxyapatite columns is presented.
D000042:Absorption; D000818:Animals; D002021:Buffers; D002417:Cattle; D004247:DNA / Q000302:isolation & purification*; D017886:Durapatite; D006882:Hydroxyapatites; D010710:Phosphates; D011506:Proteins / Q000302:isolation & purification; D012313:RNA / Q000302:isolation & purification; D013950:Thymus Gland / Q000032:analysis
D002021:Buffers; D006882:Hydroxyapatites; D010710:Phosphates; D011506:Proteins; D012313:RNA; D004247:DNA; D017886:Durapatite