Obi F O
J Chromatogr. 1986 Nov 21;369(2):321-6. doi: 10.1016/s0021-9673(00)90138-9.
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.
研究了羟基磷灰石在用于柱平衡、样品上样和柱洗涤的磷酸盐溶液中有效结合DNA的能力。结果表明，当用于柱平衡、样品上样和柱洗涤的裂解液或尿素 - 磷酸盐中的磷酸盐浓度为0.24 M时，大量DNA（高达40%）在洗涤缓冲液中被洗脱。将尿素 - 磷酸盐溶液中的磷酸盐浓度从0.24 M降低到0.15 M会导致几乎100%的结合，而在裂解液中进行类似降低则不会。本文提出了一种从羟基磷灰石柱上样和洗脱DNA的改进方法。
