Sutherland J C, Monteleone D C, Mugavero J H, Trunk J
Anal Biochem. 1987 May 1;162(2):511-20. doi: 10.1016/0003-2697(87)90427-1.
Unidirectional pulsed-field electrophoresis improves the separation of single-stranded DNA molecules longer than 20 kilobases (kb) in alkaline agarose gels compared to static-field electrophoresis. The greatest improvement in separation is for molecules longer than 100 kb. The improved resolution of long molecules with unidirectional pulsed-field electrophoresis makes possible the measurement of lower frequencies of single-strand breaks. The analytical function that relates the length and mobility of single-stranded DNA electrophoresed with a static field also applies to unidirectional pulsed field separations. Thus, the computer programs used to measure single-strand breaks are applicable to both undirectional pulsed- and static-field separations. Unidirectional pulsed-field electrophoresis also improves the separation of double-stranded DNA in neutral agarose gels. The function relating molecular length and mobility for double-stranded DNA separated by unidirectional pulsed-field electrophoresis is a superset of the function for single-stranded DNA. The coefficients of this function can be determined by iterative procedures.
与静态场电泳相比，单向脉冲场电泳可改善碱性琼脂糖凝胶中长度超过20千碱基（kb）的单链DNA分子的分离效果。对于长度超过100 kb的分子，分离效果改善最为显著。单向脉冲场电泳对长分子分辨率的提高使得测量单链断裂的较低频率成为可能。将静态场电泳的单链DNA的长度与迁移率相关联的分析函数同样适用于单向脉冲场分离。因此，用于测量单链断裂的计算机程序适用于单向脉冲场和静态场分离。单向脉冲场电泳还可改善中性琼脂糖凝胶中双链DNA的分离效果。由单向脉冲场电泳分离的双链DNA的分子长度与迁移率之间的函数关系是单链DNA函数的超集。该函数的系数可通过迭代程序确定。
