Biomedical Genomics Center: Next-Generation Sequencing Instrumentation

描述（由申请人提供）：生物研究正处于近几十年来不断改变研究的技术革命的尖端。随着“下一代”DNA测序仪的到来，单个实验室能够提出仅仅五年前需要整个机构资源的问题。新测序仪的转型方面不仅源于它们在每次运行的碱基方面的更高的总体通量，而且还源于它们允许DNA标签的数字DNA“计数”的能力。事实上，最新的下一代测序仪每次运行可计算数亿个DNA标签，与上游序列选择和制备方案相结合，有望改变基因表达、调节和表观遗传学的测量方法。这个共享仪器补助金要求这样的下一代测序仪-Illumina GA IIx。 UMN的科学家们已经为下一代测序设想了数十种有趣的应用，但目前，由于缺乏真正高性能的测序仪，这些应用受到了阻碍。他们希望使用下一代转录本计数来研究癌细胞的控制并发现调控RNA;他们希望对患者DNA进行靶向重测序，以发现导致化疗反应差异的遗传变异;他们希望使用基于序列的插入突变表征来研究致癌作用并揭示控制减数分裂基本基础的突变。一旦人们能够以合理的成本，在合理的时间范围内读取数百亿个DNA碱基，所有这些问题都成为可能。然而，如果没有这种能力，有趣的问题仍然没有答案，重要的假设也没有得到验证。 UMN的生物医学基因组学中心（BMGC）目前在UMN提供基因组服务，BMGC建议将GA IIx用于整个大学的共享使用。我们的12名科学家拥有充分利用GA IIx所需的专业知识、设施和其他资源。虽然我们目前在BMGC中运行下一代测序仪的早期模型，但在每次运行的测序读数数量和每个碱基的成本的关键指标中，这种已经使用两年的仪器不是一个可行的选择。简而言之，GAIIx将使目前无法实现的调查成为可能，因此将帮助UMN研究人员将科学更接近于治疗重要的人类疾病。

DESCRIPTION (provided by applicant): Biological research is at the cusp of a technological revolution of the type that has repeatedly transformed research in recent decades. With the arrival of "next-generation" DNA sequencers, individual laboratories are able to ask questions that just five years ago required the resources of an entire institution. The transformational aspect of the new sequencers stems not only from their higher overall throughput in terms of bases per run, but also from their ability to permit digital DNA "counting" of DNA tags. Indeed, the newest of the next-gen sequencers count hundreds of millions of DNA tags per run, which in combination with up-stream sequence selection and preparation protocols, are poised to transform measurements of gene expression, regulation, and epigenetics. This Shared Instrumentation Grant requests such a next-generation sequencer-the Illumina GA IIx. Scientists at the UMN have imagined dozens of interesting applications for next-generation sequencing, but currently, they are hampered by our lack of a truly high-powered sequencer. They would like to use next-gen counting of transcripts to study control of cancer cells and to discover regulatory RNAs; they would like to carry out targeted re-sequencing of patient DNA to discover genetic variants responsible for differences in chemotherapy response; they would like to use sequence-based characterization of insertional mutagenesis to study carcinogenesis and to uncover mutations controlling the fundamental basis of meiosis. All of these questions are made possible once one can read, at a reasonable cost, and in a reasonable time frame, tens of billions of bases of DNA. Without this capability, however, interesting questions remain unanswered, and important hypotheses untested. The UMN's Biomedical Genomics Center (BMGC) currently provides genomic services at the UMN, and the BMGC proposes to put the GA IIx into shared use for the entire university. Our twelve scientists have the expertise, facilities, and other resources required to make best use of a GA IIx. Although we presently operate an early model of next-generation sequencer in the BMGC, in the critical metric of number of sequencing reads per run and cost per base, this now two-year old instrument is not a viable option. In short, the GAIIx will enable inquiries that are simply not achievable at present, and will therefore help UMN researchers move science closer to cures for important human diseases.