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聚合酶链反应扩增寡核苷酸聚合物网络中的毛细管区带电泳:先天性肾上腺皮质增生症的案例

Capillary zone electrophoresis in polymer networks of polymerase chain reaction-amplified oligonucleotides: the case of congenital adrenal hyperplasia.

作者信息

Gelfi C, Orsi A, Righetti P G, Zanussi M, Carrera P, Ferrari M

机构信息

Faculty of Pharmacy, University of Milan, Italy.

出版信息

J Chromatogr B Biomed Appl. 1994 Jul 1;657(1):201-5. doi: 10.1016/0378-4347(94)80088-x.

Abstract

The use of capillary zone electrophoresis (CZE) in polymer networks for the analysis of an 8 bp (base pair) deletion in congenital adrenal hyperplasia was investigated. Separations were performed in Tris-borate-EDTA buffer (pH 8.3) containing 6% liquid linear polyacrylamide as a sieving dynamic matrix and 10 microM ethidium bromide for improving DNA fragment separation. Easy analysis and detection of the 127 and 135 bp amplified fragments was accomplished. The capillary column can be used for > 50 analyses before degradation and loss of resolution. The results are comparable to those obtained by gel-slab zone electrophoresis in a 12%T, 4%C polyacrylamide matrix. The sensitivity, by simple UV absorption at 254 nm, is similar to that obtained in gel slabs by dye intercalation staining.

摘要

研究了毛细管区带电泳(CZE)在聚合物网络中用于分析先天性肾上腺增生中8个碱基对(bp)缺失的情况。在含有6%液体线性聚丙烯酰胺作为筛分动态基质和10微摩尔溴化乙锭的Tris-硼酸盐-EDTA缓冲液(pH 8.3)中进行分离,以改善DNA片段分离。实现了对127和135 bp扩增片段的简便分析和检测。毛细管柱在降解和分辨率丧失之前可用于50多次分析。结果与在12%T、4%C聚丙烯酰胺基质的凝胶板区带电泳中获得的结果相当。通过在254 nm处简单的紫外吸收测得的灵敏度,与在凝胶板中通过染料嵌入染色获得的灵敏度相似。

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