Unidirectional pulsed-field electrophoresis of single- and double-stranded DNA in agarose gels: analytical expressions relating mobility and molecular length and their application in the measurement of strand breaks.
Unidirectional pulsed-field electrophoresis of single- and double-stranded DNA in agarose gels: analytical expressions relating mobility and molecular length and their application in the measurement of strand breaks.
复制标题
琼脂糖凝胶中单链和双链 DNA 的单向脉冲场电泳：与迁移率和分子长度相关的分析表达式及其在链断裂测量中的应用。
DOI：
10.1016/0003-2697(87)90427-1
复制
发表时间：
1987
影响因子：
2.9
通讯作者：
Trunk,J
中科院分区：
文献类型：
--
作者：
Sutherland,JC;Monteleone,DC;Mugavero,JH;Trunk,J
与静电场电泳法相比，单向脉冲场电泳法提高了碱性琼脂糖凝胶中大于20kb的单链DNA分子的分离能力。分离方面的最大改进是对长度超过100kb的分子。单向脉冲场电泳长分子分辨率的提高使测量单链断裂的较低频率成为可能。将单链DNA电泳法与静电场的长度和迁移率联系起来的解析函数也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既适用于单向脉冲分离，也适用于静电场分离。单向脉冲场电泳也改善了中性琼脂糖凝胶中双链DNA的分离。单向脉冲场电泳法分离的双链DNA的分子长度和迁移率相关的功能是单链DNA功能的超集。该函数的系数可以通过迭代程序来确定。
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.