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首页> 外文期刊>Chromatographia >Rapid and reproducible capillary electrophoretic separation of double-stranded DNA fragments in a simple methyl cellulosic sieving system
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Rapid and reproducible capillary electrophoretic separation of double-stranded DNA fragments in a simple methyl cellulosic sieving system

机译：在简单的甲基纤维素筛分系统中快速且可重现的毛细管电泳分离双链DNA片段

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摘要

A rapid, robust and reproducible method providing excellent separation performance and simplicity using a 0.5 % MC-4000 methyl cellulosic sieving medium in DB-1 coated capillaries has been developed. The method is suitable for qualitative comparison of DNA restriction profiles for fragments in the size range 100-1000 base pairs (bp). Efficiencies up to 8.5 million plates/m (1057 bp fragment) were recorded. Peak resolution of 6 bp (291/297 bp, 335/341 bp) and 4 bp (238/242 bp, 341/345 bp) was achieved. In addition, 1 bp partial resolution of 123/124 bp and 298/297 bp was obtained. Run-to-run (n = 15), day-to-day (n = 4), and capillary-to-capillary (n = 3) variations of 0.1-0.2 % RSD, 0.3-0.5 % RSD, and 0.1-0.3 % RSD, respectively, were observed. The MC-4000 sieving matrix was found to be better than hydroxypropyl methyl cellulose and hydroxypropyl cellulose, in terms of both performance and stability in the DB-1 coated capillaries. The efficiency and resolution in DB-WAX capillaries were inferior to those obtained in DB-1 capillaries. The commercially available DB-1 capillaries were stable for months in the sieving medium at pH 8.3 and could be regenerated to provide high efficiency after accidental current breaks. [References: 34]
机译：已开发出一种快速，耐用且可重现的方法，该方法在DB-1涂层毛细管中使用0.5％MC-4000甲基纤维素筛分介质可提供出色的分离性能和简便性。该方法适用于定性比较大小为100-1000个碱基对（bp）的片段的DNA限制性酶切图谱。记录的效率高达850万板/米（1057 bp片段）。达到了6 bp（291/297 bp，335/341 bp）和4 bp（238/242 bp，341/345 bp）的峰分辨率。另外，获得了123/124 bp和298/297 bp的1 bp部分分辨率。每次运行（n = 15），每天（n = 4）和毛细管-毛细管（n = 3）变化的RSD为0.1-0.2％，RSD为0.3-0.5％和0.1-分别观察到0.3％RSD。在DB-1涂层毛细管的性能和稳定性方面，发现MC-4000筛分基质优于羟丙基甲基纤维素和羟丙基纤维素。 DB-WAX毛细管的效率和分离度不如DB-1毛细管。市售的DB-1毛细管在pH 8.3的筛分介质中可稳定数月，并可在意外电流中断后进行再生以提供高效率。 [参考：34]

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