High Throughput Sequencer

描述（由申请人提供）：在过去十年中，大规模DNA测序显著改变了现代生物学的面貌。它的主要影响可能是产生了许多生物体的全基因组序列。这使得实验生物学在日常工作中的开展方式发生了巨大的转变，并从基因组内容的角度为理解整个生物体奠定了基础。一般来说，目前的大规模测序技术是劳动密集型的，需要大量的基础设施。试剂的每碱基成本对于任何个体研究者产生大量序列信息也是令人望而却步的。新的454生命科学测序仪使用的技术（如下所述）明显不同于目前广泛使用的ABI荧光测序仪。使用454测序仪，序列生成所需的成本、时间和劳动力大大减少。事实上，差异如此之大，以至于允许许多新的高通量应用，而这些应用对于常规测序方法是不实用的。在冷泉港实验室，一些研究人员一直在通过合作和公司提供的商业测序服务来利用454技术。还有许多问题对正在进行的项目至关重要，这些项目将在未来使用454。总体而言，该技术在以下领域对CSHL至关重要：1）比较基因组学。2)对大量患者样本进行测序，以确定有限数量的疾病候选基因中的突变（例如，在200个乳腺癌样本中搜索因基因座扩增或缺失而与疾病相关的基因外显子突变）。3)在有限数量的患者样本或组织中对基因类别进行广泛测序（例如，在大量乳腺肿瘤中调查激酶外显子的突变）。4)测定生物化学纯化的复合物、组织或生物体的小RNA含量（例如siRNA、microRNA）。5)蛋白质-核酸相互作用的全基因组测量（例如在RNP或染色质IP中）。这五个领域中的每一个都是在不同的CSHL实验室中独立出现的，每个实验室目前都在非理想的情况下进行他们的项目。由于这种类型的测序在实验室如此广泛的科学工作中变得如此重要，CSHL显然需要专用的454仪器。现场提供这种技术不仅对支持正在进行的重要研究工作至关重要，而且对鼓励以及时和具有成本效益的方式开发新项目也至关重要。

DESCRIPTION (provided by applicant): Over the last decade, large scale DNA sequencing has markedly changed the landscape of modern biology. Its major impact has probably been the generation of whole genome sequences for numerous organisms. This has created a massive shift in how experimental biology is carried out on a day-to-day basis and has built the foundation for understanding an organism as a whole, from the standpoint of its genomic content. In general, current large-scale sequencing technologies are labor intensive and require substantial infrastructure. The per-base cost of reagents is also prohibitive for any individual investigator to generate substantial sequence information. The new 454 Life Sciences sequencer uses a technology (described below) markedly different than the ABI fluorescence-based sequencers that are currently in wide use. With the 454 sequencer, the cost, time and labor required for sequence generation are greatly reduced. Indeed, the difference is so great as to permit many new high-throughput applications that are not practical with conventional sequencing methods. At Cold Spring Harbor Laboratory, a number of investigators have been making use of the 454 technology through collaborations and through a commercial sequencing service offered by the company. There are also numerous questions critical to ongoing projects that will make use of 454 in the future. Overall, the technology is critical to CSHL in the following areas: 1) Comparative genomics. 2) Sequencing of a large set of patient samples to identify mutations in a limited number of disease candidate genes (e.g. searching 200 breast cancer samples for mutations in the exons of a gene that has been linked to disease because of amplification or deletion of the locus). 3) Broad sequencing of gene classes in a limited number of patient samples or tissues (e.g. surveying kinase exons for mutations in a large set of breast tumors). 4) Determining the small RNA content (e.g. siRNAs, microRNAs) of a biochemically purified complex, a tissue or an organism. 5) Genome-wide measurements of protein-nucleic interactions (e.g. in RNP or chromatin IPs). Each of these 5 areas arose independently with in different CSHL laboratories, each of which is currently pursuing their projects under non-ideal circumstances. As this type of sequencing is becoming so central to such a wide range of scientific efforts at the Laboratory, it has become clear that CSHL requires a dedicated 454 instrument. The availability of this technology on site is essential not only to support important ongoing research efforts but also to encourage new project development in a timely and cost-effective fashion.