利用毛细管凝胶电泳对DNA限制性片段和聚合酶链反应产物进行分子大小测定。

Molecular size determinations of DNA restriction fragments and polymerase chain reaction products using capillary gel electrophoresis.

作者信息

Marino M A, Turni L A, Del Rio S A, Williams P E

机构信息

Developmental Laboratory, Armed Forces Institute of Pathology, Washington, DC 20306-6000.

出版信息

J Chromatogr A. 1994 Jul 29;676(1):185-9. doi: 10.1016/0021-9673(94)80460-5.

DOI:10.1016/0021-9673(94)80460-5

PMID:7921175

Abstract

Commercially available capillary electrophoresis (CE) systems offer advantages over traditional slab gel methodologies. The capillary format allows the use of higher voltages (225 V/cm), which results in faster migration, higher resolution and greater efficiency without excessive heating. The ability to automate the system increases the unattended sample analysis throughput. For this study, the CE system was configured with a muPAGE 3% T, 3% C polyacrylamide gel capillary with an effective length of 40 cm and microPAGE Tris-borate urea buffer system. The analysis of DNA restriction fragments and polymerase chain reaction products, with an internal standard of Boehringer Mannheim DNA marker XI were completed in less than 70 min. All samples were analyzed at 260 nm. The data establishes automated capillary gel electrophoresis as a high-resolution, reproducible method for analysis of samples under 1000 base pairs.

摘要

市售的毛细管电泳（CE）系统比传统的平板凝胶方法具有优势。毛细管形式允许使用更高的电压（225 V/cm），这可实现更快的迁移、更高的分辨率和更高的效率，且不会产生过多热量。系统自动化的能力提高了无人值守的样品分析通量。在本研究中，CE系统配置了有效长度为40 cm的muPAGE 3% T、3% C聚丙烯酰胺凝胶毛细管和microPAGE Tris-硼酸盐尿素缓冲系统。采用勃林格殷格翰DNA标记物XI作为内标，对DNA限制性片段和聚合酶链反应产物的分析在不到70分钟内完成。所有样品均在260 nm处进行分析。该数据证实了自动化毛细管凝胶电泳是一种用于分析1000个碱基对以下样品的高分辨率、可重复的方法。