CAPILLARY HYBRIDIZATION FOR IDENTIFICATION OF MUTATIONS

描述：(申请者描述)长期目标是 测量人类的突变谱，以确定原因和 导致突变、癌症和衰老的机制，以及应用 开发的突变光谱分析技术可立即应用于临床 申请。 建议开发一种明确的识别和识别的快速方法 包含突变特征的突变的测量。这一方法将 不需要对单个突变体进行分离和测序，将在 与快速、灵敏、高效的分离方法相结合 突变体称为恒变性毛细管电泳法(CDCE)。 鉴定是基于样本与一组 毛细管内先前测序的突变体，表示不同。 通过与所得到的异双链相关联的迁移率的变化。这个 这种方法的威力在于，一旦一组具有代表性的突变体 特定的序列被确定，随后的光谱解释 样品将会非常高效。 初步研究已经证明了识别个体的能力 人类组织复杂突变谱中的突变。该计划的目标是 建议包括应用于癌症相关基因的突变，如 N-ras和P53。该方法将进一步发展，用于识别和 非变性分离的双链DNA物种的测定 电泳法。

DESCRIPTION: (Applicant's Description) The long-term objective is to measure mutational spectra in humans in order to determine the causes and mechanisms leading to mutations, cancer and aging, as well as to apply the technology developed for mutational spectrometry to immediate clinical applications. It is proposed to develop a rapid method for unambiguous identification and measurement of mutants comprising mutational profiles. The approach will not require isolation and sequencing of individual mutants and will work in conjunction with the fast, sensitive and efficient method for separation of mutants called constant denaturant capillary electrophoresis (CDCE). Identification is based on DNA hybridization of a sample with a panel of previously sequenced mutants inside the capillary, Nonidentity is indicated by a change in the mobility associated with resultant heteroduplexes. The power of this approach is that once a representative set of mutants in a particular sequence is determined, interpretation of spectra in subsequent samples will be very efficient. Preliminary studies have demonstrated the ability to identify individual mutants in complex mutational spectra of human tissues. The aims of the proposal include application to cancer-associated mutations in such genes as N-ras and p53. The method will be further developed for identification and measurement of double stranded DNA species as separated by non-denaturing electrophoresis.