Acquisition of an LC Mass Spectrometer for Nucleic Acids Research

描述（由申请人提供）：本提案要求提供资金，以获得超高效液相色谱-质谱（UPLC-MS）系统，用于核酸（包括含有修饰核苷的核酸）的鉴定和结构表征。参与此请求的NIH资助的研究人员需要LC-MS/MS或LC-IMS-MS/MS功能，以实现与其资助项目相关的目标。具体地，通过这种获取将能够进行以下类型的分析：a）完整核酸的精确分子量测量; B）分离的核酸或核酸混合物的精确酶促作图; c）通过MS/MS对寡核苷酸和酶促消化混合物进行测序; d）鉴定修饰的核苷及其在核酸序列中的位置; e）使用MRM扫描的假尿苷（最丰富的修饰的核苷）的序列放置; f）寡核苷酸合成副产物和副产物的高灵敏度检测和鉴定; g）寡核苷酸：肽交联的检测;和h）修饰的RNA如转移RNA（tRNA）的从头测序。PI在质谱领域活跃了近20年，将负责所需仪器的管理监督。他将与由各参与机构的科学家组成的咨询委员会协调监督工作。它们将共同确定时间安排的优先次序，与会计人员合作确定费用回收率，并作为预测所要求设备的未来使用情况的资源。设备将存放在辛辛那提大学质谱设施中。这3,500平方米。英尺该设施于2001年进行了翻修，并配备了提供拟议分析服务所需的辅助仪器。实验室的联合主管是两个博士一级科学家。其中一名科学家将被指定为新仪器的主要操作员，预计该设施将增加一名B.S.一名高级技术员，以确保及时提供现有和新的分析服务。 公共卫生相关性：本申请中要求的UPLC-MS将允许NIH支持的研究人员访问其研究计划所需的现代仪器。这些研究人员正在解决各种与健康相关的问题，包括：抗生素耐药性，癌症，药物开发和纳米生物技术。

DESCRIPTION (provided by applicant): This proposal requests funds to acquire an ultra-high-performance liquid chromatography mass spectrometry (UPLC-MS) system intended for the identification and structural characterization of nucleic acids, including those containing modified nucleosides. The NIH-funded researchers participating in this request require LC-MS/MS or LC-IMS-MS/MS capabilities to accomplish goals associated with their funded projects. Specifically, the following types of analyses will be enabled by this acquisition: a) Accurate molecular weight measurements of intact nucleic acids; b) Accurate enzymatic mapping of isolated nucleic acids or mixtures of nucleic acids; c) Sequencing of oligonucleotides and enzymatic digestion mixtures by MS/MS; d) Identification of modified nucleosides and their placement within nucleic acid sequences; e) Sequence placement of pseudouridine, the most abundant modified nucleoside, using MRM scans; f) High sensitivity detection and identification of oligonucleotide synthetic side- and by-products; g) Detection of oligonucleotide:peptide cross-links; and h) de novo sequencing of modified RNAs, such as transfer RNAs (tRNAs). The PI, who has been active in the field of mass spectrometry for nearly 20 years, will be responsible for the managerial oversight of the requested instrumentation. He will coordinate oversight with an advisory board composed of scientists from various participating institutions. Together they will establish scheduling priorities, work with accounting personnel to establish cost-recovery rates, and will serve as a resource for projecting future use of the requested equipment. The equipment will be housed in the University of Cincinnati Mass Spectrometry Facility. This 3,500 sq. ft. facility was renovated in 2001 and is equipped with the ancillary instrumentation required for providing the proposed analytical services. The facility's co-directors are two Ph.D.-level staff scientists. One of these scientists will be designated as the primary operator of the new instrument, and it is anticipated that the facility will add a B.S.-level staff technician to ensure that existing and new analytical services are provided in a timely manner. PUBLIC HEALTH RELEVANCE: The UPLC-MS requested in this application will allow NIH-supported researchers access to modern instrumentation necessary for their research programs. These researchers are addressing a variety of health-related issues including: antibiotic resistance, cancer, drug development and nanobiotechnology.