ABI-PRISM 7700 TO ASSESS MRNA AND DNA

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，但这些方法不敏感，准确或可重复性。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，检测过程始终在PCR的线性范围内。ABI-Prism 7700还可以检测到42个副本或起始材料。不需要PCR后步骤，从而节省时间和费用。事实上，如果没有ABI-PRISM 7700的新技术，许多拟议的研究都无法进行，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的好处感到兴奋，并且这种共享仪器代表了合作的机会。主要使用者小组由来自四个学系的七名研究兴趣广泛的人士组成；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。美国国立卫生研究院资助的研究人员的项目将使用ABI-prism 7700来：(1)评估疟疾和疱疹病毒感染中CD4+ T细胞和其他选定细胞类型的细胞因子mRNA水平；(2)量化小鼠感染期间疟原虫和疱疹病毒的生物体DNA水平；(3)评估骨髓中少量前T细胞群中T细胞和淋巴细胞特异性基因的表达水平；(4)评估不同离子状态下医学上重要模型中离子转运基因的mRNA水平。(5)评估黏附分子在炎症肠组织和分选细胞中的表达；(6)确定顺式和反式作用突变对酵母HSP90基因转录的影响；(7)确定马疱疹病毒基因缺失对其他病毒基因转录的影响。ABI-Prism 7700是一种昂贵的、最先进的仪器，将最大限度地发挥美国国立卫生研究院资助的七名研究人员的研究成果。LSUMC-S管理部门热情支持这一建议，并将提供技术人员和帮助支付维修费用。该仪器将成为分子结构和动力学矿石设施的一部分；因此，已经有了维护、共享、监测和确保最佳使用这一共享仪器的程序。

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.