SpectruMedix 9610 Mutation Discovery System

SpectruMedix 9610 突变发现系统

• 批准号: 6576498
• 负责人: TERRY MAGNUSON
• 金额: $ 27.28万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-05-01 至 2004-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6576498
• 关键词: biomedical equipment purchase; gene mutation; high performance liquid chromatography; molecular genetics

DESCRIPTION (provided by applicant): The SpectruMedix SCE9610 Reveal Mutation Discovery Instrument System allows for 96 parallel analytical separations in a plate format for mutation detection and DNA polymorphism screening by lemporal gradient capillary electrophoresis (TGCE). At a temperature close to that of a variant amplicon melting temperature, the randomly reannealed combinations of homoduplexes and heteroduplexes can be separated. By comparing the electropherogram patterns of the wild-type control with experimental samples, sequence variation positives can be readily revealed. Compared with the currently used Denatured High Performance Liquid Chromatography (DHPLC) mutation detection technology, the SCE9610 offers distinctive advantages. First, unlike the DHPLC system, which is equipped with a single separation column that allows analysis of one sample at a time, separations on the SCE9610 occur simultaneously in individual capillaries. Each capillary is therefore the equivalent of a singlecolumn chromatography instrument. The SCE9610 uses one CCD detector to simultaneously record Fluorescencesignal from all capillaries with on-column laser excitation. This simple yet highly efficient optical design renders an extremely robust detection system. Most importantly, the SCE9610 configuration reduces the amount of time needed to analyze a 96-well microtiter dish from 16 hours for the current DHPLC system to less than two hours. Another limitation of the current DHPLC system is the need to establish specific temperatures for distinct types of mutations for optimal mutation detection sensitivity. The SCE9610 system circumvents this limitation by its capacity to perform sample analysis over a precisely controlled temperature gradient within a single run, eliminating the need for temperature optimization to maximize the detection sensitivity. The combination of massive multiplexing, high separation efficiency, fast analysis time, full automation and sensitive detection makes the SCE9610 a highly suitable analytical platform for high-throughput applications for many projects on the UNC-Chapel Hill campus.

描述（由申请方提供）：SpectruMeetry SCE 9610 Reveal Mutation Discovery Instrument System允许以平板形式进行96次平行分析分离，用于通过膜孔梯度毛细管电泳（TGCE）进行突变检测和DNA多态性筛查。在接近变体扩增子解链温度的温度下，可以分离同源双链体和异源双链体的随机再退火组合。通过比较野生型对照与实验样品的电泳图谱，可以容易地揭示序列变异阳性。与目前使用的变性高效液相色谱（DHPLC）突变检测技术相比，SCE9610具有明显的优势。首先，与DHPLC系统不同的是，它配备了一个单独的分离柱，允许一次分析一个样品，而SCE9610上的分离在各个毛细管中同时发生。因此，每根毛细管相当于一台色谱柱色谱仪。SCE9610使用一个CCD检测器，通过柱上激光激发，同时记录所有毛细管的荧光信号。这种简单而高效的光学设计提供了一个非常强大的检测系统。最重要的是，SCE9610配置将分析96孔微量滴定板所需的时间从当前DHPLC系统的16小时减少到不到2小时。当前DHPLC系统的另一个限制是需要为不同类型的突变建立特定的温度，以获得最佳的突变检测灵敏度。SCE9610系统能够在单次运行中通过精确控制的温度梯度进行样品分析，从而避免了这一限制，无需进行温度优化以最大限度地提高检测灵敏度。大规模多路复用、高分离效率、快速分析时间、全自动化和灵敏检测的组合使SCE9610成为北卡罗来纳大学教堂山校区许多项目高通量应用的非常合适的分析平台。