CANCER GENOTYPING WITH CAPILLARY ARRAY ELECTROPHORESIS

这部分计划项目赠款的长期目标是： 约翰霍普金斯是开发高速，大规模并行毛细管阵列 电泳装置和方法可用于有效地 癌症基因型的多重分析。这些研究将提供 可用于癌症诊断应用的技术， 导致识别与以下疾病相关的特定基因改变 癌症发展最初，毛细管阵列扫描仪能够 在不到1分钟的时间内进行多达1024次DNA扩增子电泳分离 小时将在伯克利构建，评估和优化。一个科学家 来自约翰霍普金斯将在伯克利工作，以帮助优化这一点， 设备，并学习如何运行设备和分析这些数据。 然后这个仪器将被转移到约翰霍普金斯，在那里它将被用于 癌症基因分型研究。初始系统将以1/4运行 容量装载256个样品。它将升级为领先，并运行到 1024个毛细管，因为有更多的样品和ET引物可用。自定义 ET引物将在整个工作中由技术人员合成， 伯克利与此同时，一名研究生将在伯克利工作， 毛细管阵列电泳系统的微加工 玻璃晶片和芯片扫描仪的发展。这些毛细管 阵列电泳微孔板将设计为运行96个样品， 进行分离的速度比传统的 毛细管阵列装置。样品将装载在 芯片与多头移液器和检测将提供定制 芯片扫描仪如果这些芯片成功，芯片扫描仪和芯片 将提供给约翰霍普金斯进行癌症基因分型评估 在这个补助金的第三年学习。CAE微孔板应 特别是对于实时分析过程中采集的样品 评估肿瘤进展并提供下一代 在廉价、快速和高通量癌症基因分型和诊断中 装置.

The long-term objective of this portion of the Program Project Grant with Johns Hopkins is to develop high-speed, massively parallel capillary array electrophoresis apparatus and methods that can be used for efficient multiplex analysis of cancer genotypes. These studies will provide technologies that can be used for cancer diagnostic applications and will lead to the identification of specific gene alterations associated with cancer development. Initially, a capillary array scanner capable of performing up to 1024 DNA amplicon electrophoretic separations in under 1 hour will be constructed, evaluated and optimized at Berkeley. A scientist from John Hopkins will work at Berkeley to aid in the optimization of this apparatus and to learn how to run the apparatus and analyze these data. Then this apparatus will be moved to John Hopkins where it will be used in cancer genotyping studies. The initial system will be operated at 1/4 capacity loading 256 samples. It will be upgraded to lead and run up to 1024 capillaries as more samples and ET primers become available. Custom ET primers will be synthesized throughout this work by a technician at Berkeley. In a parallel effort, a graduate student will work at Berkeley on the microfabrication of capillary array electrophoresis systems on 4" glass wafers and on the development of chip scanners. These capillary array electrophoresis microplates will be designed to run 96 samples and to perform separation that are 10-times faster than the conventional capillary array apparatus mentioned above. Samples will be loaded on the chips with multihead pipetters and detection will be provided with custom chip scanners. If these chips are successful, a chip scanner and chips will be provided to Johns Hopkins for evaluation in cancer genotyping studies in the third year of this grant. The CAE microplates should be particularly valuable for real-time analysis of samples taken during surgery to evaluate tumor progression and to provide the next generation in inexpensive fast and high through-put cancer genotyping and diagnostic devices.