ABI-PRISM 7700 TO ASSESS MRNA AND DNA

ABI-PRISM 7700 用于评估 mRNA 和 DNA

• 批准号: 2805356
• 负责人: HENRI C VAN DER HEYDE
• 金额: $ 11.88万
• 依托单位: LOUISIANA STATE UNIV HSC SHREVEPORT
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-04-15 至 2000-04-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2805356
• 关键词: DNA; biomedical equipment; biomedical equipment purchase; messenger RNA; nucleic acid quantitation /detection; nucleic acid sequence; polymerase chain reaction

The ABI-PRISM 7700 is an instrument that uses fluorescence-labeled oligonucleotides to assess mRNA levels by RT-PCR or DNA levels by PCR (termed the Taqman procedure). In other PCR methods, an end-point measurement is employed, but these methods are not as sensitive, accurate or reproducible. With the ABI-Prism, 7700, the PCR cycle number at which the fluorescence signal exceeds a threshold value is used to assess mRNA or DNA levels and consequently, the detection procedure is always in the linear range of PCR. The ABI-Prism 7700 also detects as few as 42 copies or starting material. No post PCR steps are required thereby saving time and expense. Indeed, many of the proposed studies cannot be performed without the novel technology incorporated into the ABI-PRISM 7700 because no other method can assess mRNA or DNA levels from the small amounts of sample material available. A large number of NIH-funded researchers at LSUMC are excited about the benefits of the ABI-Prism 7700, and the opportunities for the collaboration such shared instrumentation represents.. The Major User Group comprises seven individuals from four departments with a wide variety of research interests; this diversity reflects the collaborative nature of LSUMC. Each investigator has extensive experience in PCR and mRNA measurements, and thus has the expertise necessary to utilize the ABI-Prism 7700. The projects by NIH-funded investigators will use the ABI-prism 7700 to: (1) assess cytokine mRNA levels in CD4+ T cells and other selected cell-types in malaria and in Herpes virus infections, (2) quantify organism DNA level for Plasmodium and Herpes virus during infections in mice (3) assess the level of expression of T cell-and lymphoid-specific genes in minor populations of pre-T cells in the bone marrow, (4) assess mRNA levels of ion transport genes in medically important models of the different ion states, (5) assess adhesion molecule expression in tissue and sorted cells from an inflamed bowel, (6) to determine the effects of both cis- and trans-acting mutations on the transcription of HSP90 Genes in yeast; and (7) determine the effects of deletion of equine Herpesvirus genes on transcription of other viral genes. The ABI-Prism 7700 is an expensive, state-of-the-art instrument that will maximize the research efforts of a group of seven NIH-funded researchers. The LSUMC-S administration enthusiastically supports this proposal, and will provide the technical personnel and help cover maintenance costs. The instrument will be part of the Molecular Structure and Dynamics ore Facility; thus the procedures are already in place to maintain, share, monitor and ensure optimal use of this shared instrument.

ABI-PRISM 7700是一种使用荧光标记的寡核苷酸通过RT-PCR评估mRNA水平或通过PCR评估DNA水平的仪器（称为Taqman程序）。在其他PCR方法中，采用终点测量，但这些方法不那么灵敏、准确或可重复。使用ABI-Prism 7700，荧光信号超过阈值的PCR循环数用于评估mRNA或DNA水平，因此检测程序始终在PCR的线性范围内。ABI-Prism 7700还可以检测到少至42个拷贝或起始材料。无需PCR后步骤，从而节省时间和费用。事实上，如果ABI-PRISM 7700没有新技术，许多拟议的研究就无法进行，因为没有其他方法可以从少量可用的样品材料中评估mRNA或DNA水平。LSUMC的大量NIH资助的研究人员对ABI-Prism 7700的好处以及这种共享仪器所代表的合作机会感到兴奋。主要用户组由来自四个部门的七个人组成，他们具有广泛的研究兴趣;这种多样性反映了LSUMC的协作性质。每位研究者在PCR和mRNA测量方面都有丰富的经验，因此具有使用ABI-Prism 7700所需的专业知识。由NIH资助的研究人员的项目将使用ABI棱镜7700：（1）评估疟疾和疱疹病毒感染中CD 4 + T细胞和其它选定细胞类型中的细胞因子mRNA水平，（2）在小鼠感染期间定量疟原虫和疱疹病毒的生物体DNA水平（3）评估前病毒感染的少数群体中T细胞和淋巴细胞特异性基因的表达水平。骨髓中的T细胞，（4）评估不同离子状态的医学重要模型中离子转运基因的mRNA水平，（5）评估来自发炎肠的组织和分选细胞中的粘附分子表达，（6）确定顺式和反式作用突变对酵母中HSP 90基因转录的影响;（7）确定马疱疹病毒基因缺失对其它病毒基因转录的影响。ABI-Prism 7700是一种昂贵的、最先进的仪器，它将最大限度地发挥NIH资助的七名研究人员的研究成果。LSUMC-S政府热情支持这一建议，并将提供技术人员，并帮助支付维护费用。该仪器将是分子结构和动力学中心设施的一部分;因此，已经制定了程序，以维护、共享、监测和确保最佳使用这一共享仪器。