聚（环氧乙烷）-聚（环氧丙烷）-聚（环氧乙烷）三嵌段聚合物中毛细管电泳法对DNA片段的高速分离

High speed separation of DNA fragments by capillary electrophoresis in poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide) triblock polymer.

作者信息

Liang D, Chu B

机构信息

Department of Chemistry, State University of New York at Stony Brook, USA.

出版信息

Electrophoresis. 1998 Oct;19(14):2447-53. doi: 10.1002/elps.1150191416.

DOI:10.1002/elps.1150191416

PMID:9820966

Abstract

The high speed separation of DNA fragments by using a triblock copolymer, 25% w/v F127 (PEO99PPO69PEO99 with PEO and PPO denoting polyethylene oxide and polypropylene oxide, respectively) which is easy to handle and does not need coating of the quartz capillary, has been investigated. Two ways to decrease the run time are presented: one is to shorten the effective capillary length and the other to increase the electric field strength. In a short capillary, the sieving ability of the separation medium versus the initial band width, and the band width spreading as a function of distance traveled dominate the resolution; at high electric field strength, Joule heating could deteriorate the separation. By taking both effects into account, the phi X174/HaeIII digest could be separated within 100 s by using an 8 mm effective length, 50 microns diameter capillary operating at 300 V/cm.

摘要

研究了使用三嵌段共聚物（25% w/v F127，即PEO99PPO69PEO99，其中PEO和PPO分别表示聚环氧乙烷和聚环氧丙烷）对DNA片段进行高速分离的方法，该共聚物易于操作，且无需对石英毛细管进行涂层处理。文中提出了两种减少运行时间的方法：一种是缩短有效毛细管长度，另一种是提高电场强度。在短毛细管中，分离介质对初始带宽的筛分能力以及带宽随迁移距离的扩展主导着分辨率；在高电场强度下，焦耳热会使分离效果变差。综合考虑这两种效应，通过使用有效长度为8 mm、内径为50微米的毛细管，在300 V/cm的电场强度下操作，phi X174/HaeIII酶切片段可在100 s内分离。