Cech T R, Reisler F, Hearst J E
Biochemistry. 1976 May 4;15(9):1865-73. doi: 10.1021/bi00654a012.
A new technique--partial denaturation of DNA in equilibrium CsCl density gradients at pH 11.4--is used to determine the distribution of intermediate states in the melting of mouse DNA. When the technique is applied in the preparative ultracentrifuge, the DNA is fractionated according to stability. Neutralization of the partially denatured fractions results in the recovery of most of the DNA in its native form. The individual fractions are more homogeneous than the total DNA: they have decreased density heterogeneity (smaller band widths), neutral CsCl buoyant densities that differ from the average, and more homogeneous melting profiles with melting temperatures that differ from the average.
一种新技术——在pH 11.4的平衡CsCl密度梯度中对DNA进行部分变性——被用于确定小鼠DNA解链过程中中间状态的分布。当该技术应用于制备型超速离心机时,DNA会根据稳定性进行分级分离。对部分变性级分进行中和后,大部分DNA会以天然形式回收。各个级分比总DNA更均一:它们的密度异质性降低(带宽更小),中性CsCl浮力密度不同于平均值,并且具有与平均值不同的解链温度的更均一的解链曲线。