The goal of this project is the development of a novel type of multi-capillary container (carrier) for electrophoretic DNA separation - the monolithic multi- capillary array (MMCA). This linear array has a specially designed patented architecture and is fabricated with a proprietary technology. It has a shape of a 50-60 cm long monolith glass ribbon with capillary channels running the length of the ribbon. The main goal of Phase I of the project is the demonstration of feasibility of using MMCA as a carrier for electrophoretic DNA sequencing. To achieve this goal we will perform the following: * Design and develop modules and tools necessary to carry out DNA sequencing in a 12- and 32-channel MMCA * Carry out test sequencing runs on clinical DNA sequencing. After successful completion of Phase I, the goal of Phase II will be incorporation of MMCA in the DNA sequencing instrument for a high resolution identification of mutations in mixed DNA populations, that is currently being developed by our research group. In Phase II we also plan a broad study of DNA sequencing in MMCA. This includes the analysis of clinical tumor samples with mutant p53 gene, addressing such issues as mutation detection, heterozygote determination, as well as the sensitivity and mutation resolution in mixed populations of DNA molecules. The key advantages of MMCA's are rooted in their excellent physical parameters for capillary electrophoresis and fluorescent detection. The MMCA's are made of glass of any desired length. With the separation length easily in the range of 50-60 cm, the inner diameter of capillaries in the MMCA can be designed anywhere between 10 mu m and 100 mu m with inter-capillary distance of 50-100 mu m. The superior optical properties of MMCA's combined with the ease of installation and replacement offers a new generation of high-resolution compact inexpensive multi-capillary carriers for DNA sequencing.
The proposed novel method of the in-capillary densification of the injected DNA sample plug will find application in the DNA sequencing using capillary electrophoresis. Its most important significance is the increase of the read length of the sequencing run. This will reduce the labor intensive part of the process. The method will also lead to a lesser cost of the sequencing equipment and genetic materials.
