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使用电喷雾扫描迁移率粒度分析仪进行DNA分析。

DNA analysis using an electrospray scanning mobility particle sizer.

作者信息

Mouradian S, Skogen J W, Dorman F D, Zarrin F, Kaufman S L, Smith L M

机构信息

Department of Chemistry, University of Wisconsin, Madison 53706-1396, USA.

出版信息

Anal Chem. 1997 Mar 1;69(5):919-25. doi: 10.1021/ac960785k.

Abstract

A scanning mobility particle sizer (SMPS) allows size separation of gas phase particles according to their electrophoretic mobilities. The addition of an electrospray source (ES) recently allowed extension of SMPS analysis to the macromolecular range. We demonstrate here the application of ES-SMPS to nucleic acids analysis. Single- and double-stranded DNA molecules ranging from 6.1 kDa (single-stranded DNA 20 nucleotides in length) to 300 kDa (500 base-pair double-stranded DNA) were separated and detected by ES-SMPS at the picomole to femtomole levels. The measured electrophoretic mobility diameters were found to correlate with the analytes' molecular weights, while the peak areas could yield quantitative information. No fragmentation of DNA was observed under the conditions employed. Different apparent densities were observed for single-stranded and double-stranded DNAs, showing a different behavior for each type of biomolecule. The total analysis time was about 3 min/spectrum. Further optimization of ES-SMPS is expected to make it a fast and sensitive technique for biopolymer characterization.

摘要

扫描迁移率粒径分析仪(SMPS)可根据气相颗粒的电泳迁移率对其进行尺寸分离。最近,电喷雾源(ES)的加入使得SMPS分析能够扩展到高分子范围。我们在此展示了ES-SMPS在核酸分析中的应用。通过ES-SMPS在皮摩尔至飞摩尔水平上分离并检测了分子量范围从6.1 kDa(长度为20个核苷酸的单链DNA)到300 kDa(500个碱基对的双链DNA)的单链和双链DNA分子。发现所测量的电泳迁移率直径与分析物的分子量相关,而峰面积可提供定量信息。在所采用的条件下未观察到DNA的断裂。单链和双链DNA表现出不同的表观密度,表明每种生物分子类型具有不同的行为。每次光谱的总分析时间约为3分钟。预计对ES-SMPS的进一步优化将使其成为一种用于生物聚合物表征的快速且灵敏的技术。

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