Use of secondary pulsed field gel electrophoresis in separation of large DNA.
Use of secondary pulsed field gel electrophoresis in separation of large DNA.
复制标题
使用二次脉冲场凝胶电泳分离大 DNA。
DOI：
10.1006/abio.1995.1009
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发表时间：
1995
期刊：
影响因子：
--
通讯作者：
Birren,BW
中科院分区：
文献类型：
--
作者：
Lai,E;Birren,BW
据报道，二次脉冲场凝胶（SPFG）电泳可以显著提高分离大DNA分子的速度而不降低分辨率（Zhang，T.是的，史密斯角，澳-地L.，和Canter，C. R.（1991）Nucleic Acids Res. 19，1291-1296）。然而，我们试图复制以前的SPFG条件没有成功。因此，我们试图更精确地定义二次脉冲对分离大DNA的影响，并确定该技术在分离高达1100 kb的分子中的价值。在这里，我们报告的两个关键SPFG参数，即二次脉冲的频率和持续时间的迁移和分辨率的DNA在SPFG。我们发现，分离的大小范围由主脉冲和次级脉冲的持续时间的总和决定。在最佳条件下，可以在不损失分辨率的情况下实现速度增加25-70%。
It has been reported that secondary pulsed field gel (SPFG) electrophoresis can dramatically increase the speed of separation of large DNA molecules without a decrease in resolution (Zhang, T. Y., Smith, C. L., and Canter, C. R. (1991) Nucleic Acids Res. 19, 1291-1296). However, our attempts to duplicate previous SPFG conditions were unsuccessful. We therefore sought to more precisely define the effects of secondary pulsing on the separation of large DNA and to determine the value of the technique in separating molecules up to 1100 kb. Here we report on two of the key SPFG parameters, namely the frequency and duration of the secondary pulse on the migration and resolution of DNA in SPFG. We found that the size range of separation is determined by the sum of the duration of the primary and secondary pulses. Under optimal conditions, a 25-70% increase in velocity can be achieved without loss in resolution.