Belgrader P, Devaney J M, Del Rio S A, Turner K A, Weaver K R, Marino M A
Advanced Technologies Development Branch, Armed Forces DNA Identification Laboratory, Armed Forces Institute of Pathology, Rockville, MD 20850, USA.
J Chromatogr B Biomed Appl. 1996 Aug 9;683(1):109-14. doi: 10.1016/0378-4347(96)00165-x.
The analysis of crude polymerase chain reaction (PCR) products by capillary electrophoresis (CE) is often compromised due to the presence of a high concentration of salt. Salt interferes with the electrokinetic injection and induces localized heating within the column; hence, PCR products must be desalted or cleaned-up prior to CE analysis. A variety of commercial clean-up systems are available that have been traditionally used to prepare PCR products for cloning, sequencing and digestion with restriction enzymes. These systems were tested for their effectiveness in preparing PCR products for CE analysis and were evaluated based on CE resolution, salt removal, DNA recovery, processing time and cost. One particularly effective clean-up system, membrane dialysis, was automated using a robotic workstation.
由于存在高浓度盐，通过毛细管电泳（CE）对粗聚合酶链反应（PCR）产物进行分析常常受到影响。盐会干扰电动进样并在柱内引起局部加热；因此，在进行CE分析之前，必须对PCR产物进行脱盐或纯化处理。有多种市售的纯化系统，传统上用于制备用于克隆、测序和用限制性内切酶消化的PCR产物。测试了这些系统在制备用于CE分析的PCR产物方面的有效性，并根据CE分辨率、盐去除、DNA回收率、处理时间和成本进行了评估。一种特别有效的纯化系统——膜透析，使用机器人工作站实现了自动化。
