24-Capillary Reveal Mutation Discovery System

描述（由申请方提供）：使用SpectuMeetings Reveal-2410 Mutation Discovery System进行的温度梯度毛细管电泳（TGCE）将加速生物表型和发育过程相关DNA序列的鉴定。通过将温度梯度电泳与毛细管电泳相结合，可以基于解链温度快速鉴定核苷酸序列中具有错配的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%的效率检测大小和核酸序列不同的多个片段中的错配。这是一个主要的优势，其他方法来确定功能性核苷酸序列的变化。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可用作标准核苷酸测序仪来鉴定核苷酸变化。由于基于噬菌体的大肠杆菌的广泛使用，也需要增加核苷酸测序。大肠杆菌同源重组，用于快速修饰基因组DNA的大片段，以产生用于敲除和转基因小鼠的构建体。所要求的仪器将有利于NIH资助的研究计划，在遗传控制神经髓鞘的雪旺细胞，在功能基因组学的易感性朊病毒疾病，在分子马达的作用，在听觉系统和先天性耳聋的发育生物学。除了该技术显示200至500 bp片段中的单碱基变化（如ENU诱导的同基因突变）的能力外，新的用途（如QTL潜在的候选基因排序）可能证明是富有成效的。Spectrumedix Reveal 2410不仅将加速McLaughlin研究所目前的研究，还有助于招募神经遗传学领域的新生物医学研究人员。

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.