24-Capillary Reveal Mutation Discovery System

24-毛细管揭示突变发现系统

• 批准号: 6578473
• 负责人: George A. Carlson
• 金额: $ 9.44万
• 依托单位: MC LAUGHLIN RESEARCH INSTITUTE
• 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/6578473
• 关键词: biomedical equipment purchase; capillary electrophoresis; gene mutation; molecular genetics; nucleic acid quantitation /detection

DESCRIPTION (provided by applicant): Temperature gradient capillary electrophoresis (TGCE) using the SpectuMedix Reveal-2410 Mutation Discovery System will accelerate identification of the DNA sequence underlying biological phenotypes and developmental processes. By combining temperature gradient electrophoresis with capillary electrophoresis it is possible to rapidly identify DNA fragments with mismatches in nucleotide sequence based on the melting temperature. By running sufficiently broad temperature gradients, mismatches in multiple fragments differing in size and nucleic acid sequence can be detected at efficiencies above 95% in a single run. This is a major advantage over other approaches to identify functional nucleotide sequence changes. Rather than sequencing an entire candidate gene, the Mutation Discovery System prioritizes the exon or region of DNA with mismatch between mutant and wild-type for nucleotide sequencing. The Reveal-2410 can then be used as a standard nucleotide sequencer to identify the nucleotide changes. Increased nucleotide sequencing is also needed due to expanded use of phage-based E. coli homologous recombination for rapid modification of large segments of genomic DNA for generation of constructs for knockout and transgenic mice. The instrument requested will benefit NIH funded research programs in the genetic control of rnyelination by Schwann cells, in the functional genomics of susceptibility to prion diseases, in the role of molecular motors in hearing, and in the developmental biology of the auditory system and congenital deafness. In addition to the demonstrated ability of this technology to reveal single base changes in fragments between 200 to 500 bp, such as isogenic mutations induced by ENU, novel uses, such ranking candidate genes underlying QTL, are likely to prove fruitful. The Spectrumedix Reveal 2410 will not only accelerate current research at McLaughlin Research Institute, but help in recruiting new biomedical researchers in the field of neurogenetics.

描述（由申请人提供）：使用 SpectuMedix Reveal-2410 突变发现系统的温度梯度毛细管电泳 (TGCE) 将加速生物表型和发育过程中 DNA 序列的识别。通过将温度梯度电泳与毛细管电泳相结合，可以根据解链温度快速识别核苷酸序列中存在错配的 DNA 片段。通过运行足够宽的温度梯度，可以在单次运行中以高于 95% 的效率检测不同大小和核酸序列的多个片段中的错配。与其他识别功能性核苷酸序列变化的方法相比，这是一个主要优势。突变发现系统不是对整个候选基因进行测序，而是优先考虑突变体和野生型之间不匹配的 DNA 外显子或区域进行核苷酸测序。 Reveal-2410 然后可以用作标准核苷酸测序仪来识别核苷酸变化。由于基于噬菌体的大肠杆菌同源重组的广泛使用，可以快速修饰大片段基因组 DNA，从而生成用于敲除和转基因小鼠的构建体，因此还需要增加核苷酸测序。所需的仪器将有利于美国国立卫生研究院资助的研究项目，包括雪旺细胞髓鞘形成的遗传控制、朊病毒疾病易感性的功能基因组学、分子马达在听力中的作用以及听觉系统和先天性耳聋的发育生物学。除了该技术已证明能够揭示 200 至 500 bp 片段中的单碱基变化（例如 ENU 诱导的同基因突变）之外，新用途（例如 QTL 下的候选基因排名）也可能会取得丰硕成果。 Spectrumedix Reveal 2410 不仅将加速 McLaughlin 研究所当前的研究，而且有助于招募神经遗传学领域的新生物医学研究人员。