CAPILLARY HYBRIDIZATION FOR IDENTIFICATION OF MUTATIONS

描述：（申请人的描述）长期目标是 测量人类的突变谱，以确定原因， 导致突变、癌症和衰老的机制，以及应用 为突变光谱法开发的技术可立即用于临床 应用. 建议开发一种快速无模糊识别方法， 包含突变谱的突变体的测量。 该办法将 不需要单独的突变体的分离和测序， 结合快速、灵敏和有效的分离方法， 突变体称为恒定变性毛细管电泳（CDCE）。 鉴定是基于样品与一组 毛细管内先前测序的突变体，指示不一致性 通过与所得异源双链体相关的迁移率的变化。 的 这种方法的力量在于，一旦一组代表性的突变体在一个 确定特定序列，随后对光谱进行解释， 样品将非常有效。 初步研究表明， 人类组织复杂突变谱中的突变体。 的目标 建议包括应用于这样基因中的癌症相关突变， N-ras和p53。 该方法将进一步发展， 通过非变性分离的双链DNA物质的测量 电泳

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.