Temperature Gradient Hybridizer and Real-time Imager for DNA Microarrays

DNA 微阵列温度梯度杂交仪和实时成像仪

• 批准号: 7405099
• 负责人: George W Jackson
• 金额: $ 11.19万
• 依托单位: BIOTEX, INC.
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-02-01 至 2010-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7405099
• 关键词: Address; Agreement; Binding; Biological; Biological Assay; Compatible; Competitive Binding; Condition; DNA Microarray Chip; DNA Microarray format; DNA Probes; Data; Data Quality; Detection; Development; Devices; Elements; Engineering; Equipment; Fluorescence; Gene Expression; Genetic; Genotype; Image; Investigation; Kinetics; Label; Laboratories; Length; Location; Manufacturer Name; Marketing; Measurement; Metric; Microscope; Modeling; Molecular Biology; Nucleic Acid Hybridization; Nucleic Acids; Numbers; Oligonucleotide Probes; Oligonucleotides; Phase; Pliability; Polymerase Chain Reaction; Protocols documentation; Range; Reaction; Real-Time Systems; Reproducibility; Research Project Grants; Sampling; Sensitivity and Specificity; Slide; Specificity; Spottings; Standards of Weights and Measures; Surface; System; Techniques; Temperature; Testing; Time; Work; base; biological research; design; engineering design; improved; instrument; melting; microbial; novel; prototype; research study; tool

DESCRIPTION (provided by applicant): Because of the large amount of genetic information which can be rapidly accessed, DNA microarrays have become indispensable tools for biological investigations. Despite their utility, however, a number of experimental difficulties have resulted in widely varying data quality between and even within the same laboratories. One of the more difficult challenges is that DNA probes of the same length can have widely varying hybridization or melting temperatures (Tm's). Surprisingly, although temperature gradient PCR devices exist for rapid optimization of reaction conditions, no such commercial devices exist for microarrays. The result is that suboptimal conditions are used for what are presently some of the most expensive experiments in molecular biology. This project focuses on the development of a hardware system for producing optimal "temperature addresses" for every probe included in an arbitrary microarray design. This improvement will dramatically improve the accuracy and reproducibility of nucleic acid microarray data. By improving specificity of binding, the limit of detection for rare sequences in a sample should also be improved. Finally, the system will greatly increase flexibility regarding the ability to include sequences which would have been previously discarded from probe sets. As microarrays continue to find expanding applications in biological research, the developed system will represent an important advance in the field and should have considerable commercial attractiveness. In Phase II, we will integrate the temperature gradient hybridization hardware with a CCD-based imaging system for real-time image acquisition. Real-time imaging will allow kinetics to be followed as well as non-isothermal (time-varying gradients in this case) data to be acquired such as annealing kinetics, fluorescence melting curves, competitive binding assays, and "on-chip" PCR, for example. By the end of Phase II, we will able to market our well-characterized equipment as well as the proof- of-principle data to manufacturers of microarray imaging and support equipment. DNA microarrays have become an indispensable tool for biological research, however a number of technical challenges have hampered their reliable use. This project focuses on developing the equipment for providing the optimal hybridization conditions for every probe on a microarray, as well as for imaging the surface reactions on the array in real-time. The engineering advances which will result will allow increased flexibility in microarray design for a variety of genotyping applications including gene expression analysis, chromosomal accessibility, sequencing by hybridization, and microbial identification.

描述（申请人提供）：由于大量的遗传信息可以快速获取，DNA微阵列已经成为生物研究不可缺少的工具。然而，尽管它们很实用，但一些实验上的困难导致了在同一实验室之间甚至在同一实验室内部的数据质量差别很大。更困难的挑战之一是相同长度的DNA探针可以有很大差异的杂交或熔化温度（Tm）。令人惊讶的是，虽然存在温度梯度PCR装置用于快速优化反应条件，但没有这种用于微阵列的商用设备。其结果是，次优条件被用于目前分子生物学中一些最昂贵的实验。