DNA capillary electrophoresis using poly(vinyl alcohol). II. Separation media.

DNA capillary electrophoresis using poly(vinyl alcohol). II. Separation media.
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使用聚(乙烯醇)进行 DNA 毛细管电泳。

DOI:
10.1002/elps.200305547
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发表时间:
2003
期刊:
影响因子:
2.9
通讯作者:
Chu,Benjamin
Chu,Benjamin
中科院分区:
生物学3区
文献类型:
--
作者:
Moritani,Tohei;Yoon,Kyunghwan;Chu,Benjamin

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我们代表了使用各种聚乙烯醇(PVAL)相关聚合物作为分离介质的DNA毛细管电泳的第一次广泛研究。作为分离介质,PVAL均聚物显示出较差的分辨率,可能是由于其非常强的氢键特性,导致广泛的自聚集。另一方面,聚(乙烯醇-共-乙酸乙烯酯)和聚(乙烯醇-共-1-乙烯基-2-吡咯烷酮)(PVAL-VP),聚合度均为10.3 ×103,已被发现可以提供具有良好分辨率的优异电泳图,用于分析双链(ds)DNA。PVAL-VP在高pH和低pH区域具有水解稳定性,在中性和碱性条件下也产生了良好的单链(ss)DNA电泳图,尽管为了提高DNA测序分析所需的分辨率,有必要进行进一步的研究。在碱性条件下获得的分离显示出显著更短的保留时间,是目前商业分离介质的三分之一,这是由于DNA链中磷酸基团的更高电离。
We represent the first extensive study on DNA capillary electrophoresis using various poly(vinyl alcohol) (PVAL)‐related polymers as separation media. As a separation medium, PVAL homopolymer has shown poor resolutions probably due to its very strong hydrogen‐bonding characteristics, resulting in extensive self‐aggregation. On the other hand, poly(vinyl alcohol‐co‐vinyl acetate) and poly(vinyl alcohol‐co‐1‐vinyl‐2‐pyrrolidone) (PVAL‐VP), both with a degree of polymerization of ∼3.0×103, have been found to give excellent electropherograms with good resolutions for the analysis of double‐stranded (ds)DNA. PVAL‐VP, with hydrolytic stability in high and low pH regions, has also yielded fair electropherograms for single‐stranded (ss)DNA under neutral and alkaline conditions, although further investigation is essential in order to increase the resolutions necessary for DNA sequencing analysis. The separations obtained under alkaline conditions have shown significantly shorter retention times, one‐third of that for the current commercial separation media, due to the higher ionization of phosphate groups in the DNA chains.
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