WHOLE GENOME AMPLIFICATION FOR RAPID LOW COST GENOTYPING

用于快速低成本基因分型的全基因组扩增

• 批准号: 7291107
• 负责人: Roger Sender Lasken
• 金额: $ 36.72万
• 依托单位: J. CRAIG VENTER INSTITUTE, INC.
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-05-12 至 2008-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7291107
• 关键词: Address; Adoption; Affect; Archives; B-Lymphocytes; Biological; Biological Assay; Biological Models; Blood; Bypass; Cell Count; Cells; Cheek structure; Chromosome Mapping; Clinical; Collection; DNA; DNA amplification; DNA biosynthesis; DNA chemical synthesis; DNA purification; Development; Enzymatic Biochemistry; Genetic Heterogeneity; Genome; Genomics; Genotype; Goals; Haplotypes; Human; Human Genome Project; Individual; Institutes; Joints; Lymphocyte; Measures; Methods; Microfluidics; Miniaturization; Molecular; Numbers; Partner in relationship; Pharmacologic Substance; Polymerase; Preimplantation Diagnosis; Preparation; Process; Production; Protocols documentation; Range; Reaction; Research; Research Personnel; Sampling; Single Nucleotide Polymorphism; Specimen; Swab; Technology; Vision; base; cost; embryo stage 2; genetic analysis; human disease; improved; new technology; programs; reaction rate; sperm cell; trend; tumor

DESCRIPTION (provided by applicant): Whole genome amplification by the multiple displacement amplification (MDA) reaction has the potential to transform the way that DNA is prepared for genotyping assays. The long-term goal is to develop a low cost and rapid process for amplifying genomic DNA directly from minute clinical specimens or stored DNA samples. DNA amplified from blood and buccal swab samples will be ready for use in SNP assays, bypassing conventional bead or cartrige based DNA purification steps. New research applications of MDA will be investigated based on its ability to amplify DNA from single cells. The specific aims are to: 1. Optimize the MDA reaction enzymology. Molecular mechanisms will be investigated for the "hyperbranching" process that underlies exponential amplification. Reaction intermediates will be evaluated for the steps of template denaturation, random priming, and strand displacement DNA synthesis. Parameters will be defined and measures that affect reaction rate, DNA yield, amplification bias, and fidelity of sequence replication. 2. Develop a low cost, high throughput DNA sample preparation process for use in SNP genotyping. An integrated process will deliver DNA from blood and buccal swab samples directly into genotyping assays. Miniaturization and microfluidic methods will add to throughput and cost reduction. Processes will be validated for DNA preparation from archives of biological samples and rare DNA collections to increase access to a wider range of researchers. 3. Demonstrate methods for whole genome amplification from single cells. A) Single sperm will be used as a means to obtain haplotypes. B) Cells isolated from human tumors will be assessed for genetic heterogeneity in the transformation process. C) Individual lymphocytes will be used as a model system to prove feasibility of doing preimplantation genetic diagnosis from single blastomeres.

描述（由申请人提供）：通过多重位移扩增（MDA）反应的全基因组扩增有可能改变DNA用于基因分型分析的制备方式。长期目标是开发一种低成本和快速的方法，直接从微小的临床标本或储存的DNA样本中扩增基因组DNA。从血液和口腔拭子样本中扩增的DNA将准备好用于SNP分析，绕过传统的基于头或墨盒的DNA纯化步骤。基于其扩增单细胞DNA的能力，将研究MDA的新研究应用。具体目标是：