一种新型凝胶基质用于DNA片段分离:平板凝胶电泳与毛细管电泳的比较研究。

B A Siles, G B Collier, D J Reeder, W E May
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引用次数: 0

摘要

TreviGel-500是一种含有AgaCryl的新型多糖基质,可作为平板凝胶电泳的粉末,目前正应用于毛细管电泳中DNA片段的分离。与平板凝胶电泳相比,毛细管模式允许使用一到两个数量级的基质质量分数和大约五到六个数量级的样品数量,以在100到2000个碱基对大小范围内实现DNA片段的最佳分离。在毛细管模式下,这种新的分离基质形成了一种半刚性凝胶,相对于其他筛分聚合物溶液,这种凝胶对1,000至7,000碱基对大小范围内的DNA片段具有更高的选择性。此外,该基质具有比丙烯酰胺毒性低的优点。比较了在平板凝胶电泳和毛细管电泳中使用该基质分离DNA片段的缓冲液中基质的质量分数、缓冲液组成和样品装载或注射参数。
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The use of a new gel matrix for the separation of DNA fragments: a comparison study between slab gel electrophoresis and capillary electrophoresis.

TreviGel-500, a new polysaccharide matrix containing AgaCryl, commercially available as a powder for slab gel electrophoresis, is now being applied to the separation of DNA fragments in capillary electrophoresis. The capillary mode allows the use of one to two orders of magnitude lower mass fractions of matrix and approximately five to six orders of magnitude lower sample quantities than the slab gel electrophoresis counterpart for optimal separation of DNA fragments in the 100 to 2,000 base pair size range. In the capillary mode, this new separation matrix forms a semi-rigid gel that demonstrates enhanced selectivity for DNA fragments in the 1,000 to 7,000 base pair size range relative to alternative size-sieving polymer solutions. In addition, this matrix offers the advantages of lower toxicity than acrylamide. Comparisons are drawn between the use of this matrix in both slab gel electrophoresis and capillary electrophoresis for the separation of DNA fragments with respect to the mass fraction of the matrix in buffer, the buffer composition and sample loading or injection parameters.

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Discontinuous electrophoresis revisited: a review of the process. The evaluation of fast purification methods for preparing polymerase chain reaction (PCR) products for capillary electrophoresis analysis. The use of a new gel matrix for the separation of DNA fragments: a comparison study between slab gel electrophoresis and capillary electrophoresis. DNA sequencing by capillary electrophoresis with a hydroxyethylcellulose sieving buffer. The distribution of F13A subtypes in four populations using agarose isoelectric focusing and Western Blot detection.
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