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Study on Detection Methods of Gene Mutation and Their Clinical Applications

Author: ZhaoChunXia
Tutor: XuGuoWang
School: Dalian Institute of Chemical Physics
Course: Analytical Chemistry
Keywords: Genetic polymorphism Sequence variation Detection methods Clinical application
CLC: R346
Type: PhD thesis
Year: 2004
Downloads: 232
Quote: 0
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Abstract


In recent years, the organisms genome sequence variation and polymorphism studies have become the focus of attention. The technology is still not mature enough the traditional sequence variation and polymorphism detection technology is far from meeting the needs now, and be able to bulk, cheap identification and detection of sequence variation. Therefore, the establishment of an efficient, accurate and suitable for clinical application of sequence variation detection technology platform is extremely important. This paper first prepared a suitable genome sequence variation and polymorphism detection efficiency, economic capillary electrophoresis (CE) sieving medium --- short-chain linear polyacrylamide (LPA) and capillary coated column, and small fragment DNA molecule as a target gene, and its migration behavior in the short-chain LPA sieving medium, polymer characteristic DNA isolation, further validation Ogston model and polymer semidilute solution clew contraction theory on the reasonableness of the CE-DNA screening system to predict. For the establishment of a capillary electrophoresis platform for rapid screening and detection of genomic sequence variations and polymorphisms, DNA screening system of economic, serialization, loyal, paper use of home-made short-chain LPA sieving medium, type in ABI310 The genetic analyzer on a series of homemade LPA sequence polymorphism detection methods: 1) the establishment of a rapid detection of known single nucleotide polymorphisms (SNPs) in the genome and genetic typing the SNaPshot method; 2) single strand conformation polymorphism for rapid screening of unknown gene sequence variations, single-strand conformation polymorphism / heterozygosity analysis, the constant degeneration capillary electrophoresis detection method. Investigated sieving medium in each of the above-described method, the separation temperature, voltage, additives, etc. was separated mutual dependencies between its impact on the separation. Under the conditions of each optimization with homemade short-chain LPA sieving medium and commercialization sieving medium compared, the results show that better separation performance homemade LPA sieving medium and shorter analysis time and lower detection costs, and more suitable for large-scale analysis and clinical application. Useful to verify the above method, the platform is used in actual clinical samples. First, sequence variations and polymorphisms rapid screening of a tumor suppressor gene (cancer) tumor samples discussed above the crowd tumor development pathway and cancer risk. Second, the 16S rRNA gene and 16S-23S rRNA region genes belonging to 14 270 are 34 kinds of common clinical pathogens rapid analysis, preliminary clinical common pathogens double SSCP, and RFLP standard database and test results with machine code combine to make complex data simple and laid the foundation for the early clinical application of the law.

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CLC: > Medicine, health > Basic Medical > Human biochemistry, molecular biology
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