MONOLITHIC MULTI-CAPILLARY ARRAYS TO SEQUENCE DNA

本项目的目标是开发一种用于DNA电泳分离的新型多毛细管容器(载体)--整体式多毛细管阵列(MMCA)。这种线性阵列具有特别设计的专利架构，并采用专有技术制造。它具有50-60厘米长的整体玻璃带的形状，毛细管通道贯穿带子的长度。该项目第一阶段的主要目标是论证使用MMCA作为DNA电泳测序载体的可行性。为了实现这一目标，我们将执行以下工作：*设计和开发在12通道和32通道MMCA中进行DNA测序所需的模块和工具*在临床DNA测序上进行测序测试运行。在成功完成第一阶段后，第二阶段的目标将是将MMCA纳入DNA测序仪器，以高分辨率识别混合DNA群体中的突变，这是我们的研究小组目前正在开发的。在第二阶段，我们还计划在MMCA中进行广泛的DNA测序研究。这包括分析带有突变型p53基因的临床肿瘤样本，解决突变检测、杂合子确定以及DNA分子混合群体中的敏感性和突变分辨率等问题。MMCA的主要优势在于其用于毛细管电泳法和荧光检测法的优良物理参数。MMCA是由任意长度的玻璃制成的。MMCA的分离长度在50-60厘米范围内，毛细管内径可在10-100微米之间设计，毛细管间距离为50-100微米。MMCA优越的光学性能与易于安装和更换相结合，为DNA测序提供了新一代高分辨率、紧凑、廉价的多毛细管载体。建议的商业应用：建议的毛细管内致密DNA样品塞的新方法将在使用毛细管电泳法进行DNA测序中得到应用。其最重要的意义是增加了测序运行的读取长度。这将减少这一过程中的劳动密集型部分。该方法还将导致测序设备和遗传物质的成本更低。

The goal of this project is the development of a novel type of multi-capillary container (carrier) for electrophoretic DNA separation - the monolithic multi- capillary array (MMCA). This linear array has a specially designed patented architecture and is fabricated with a proprietary technology. It has a shape of a 50-60 cm long monolith glass ribbon with capillary channels running the length of the ribbon. The main goal of Phase I of the project is the demonstration of feasibility of using MMCA as a carrier for electrophoretic DNA sequencing. To achieve this goal we will perform the following: * Design and develop modules and tools necessary to carry out DNA sequencing in a 12- and 32-channel MMCA * Carry out test sequencing runs on clinical DNA sequencing. After successful completion of Phase I, the goal of Phase II will be incorporation of MMCA in the DNA sequencing instrument for a high resolution identification of mutations in mixed DNA populations, that is currently being developed by our research group. In Phase II we also plan a broad study of DNA sequencing in MMCA. This includes the analysis of clinical tumor samples with mutant p53 gene, addressing such issues as mutation detection, heterozygote determination, as well as the sensitivity and mutation resolution in mixed populations of DNA molecules. The key advantages of MMCA's are rooted in their excellent physical parameters for capillary electrophoresis and fluorescent detection. The MMCA's are made of glass of any desired length. With the separation length easily in the range of 50-60 cm, the inner diameter of capillaries in the MMCA can be designed anywhere between 10 mu m and 100 mu m with inter-capillary distance of 50-100 mu m. The superior optical properties of MMCA's combined with the ease of installation and replacement offers a new generation of high-resolution compact inexpensive multi-capillary carriers for DNA sequencing. PROPOSED COMMERCIAL APPLICATIONS: The proposed novel method of the in-capillary densification of the injected DNA sample plug will find application in the DNA sequencing using capillary electrophoresis. Its most important significance is the increase of the read length of the sequencing run. This will reduce the labor intensive part of the process. The method will also lead to a lesser cost of the sequencing equipment and genetic materials.