Hybridization-Assisted Nanopore Sequencing

描述（由申请人提供）：技术发展的基本原理将使极便宜，高速测序是很好的建立。其中最主要的是个性化医疗的实现。目前正在开发的几种技术有望显著降低人类基因组测序的成本。然而，目前尚不清楚的是，这些技术是否足以使全基因组测序成为常规的临床工具。此外，那些在这个成本参数上最有前途的技术，看起来似乎将面临诸如读取长度等性能特征方面的困难。基于纳米孔的测序技术有望廉价、快速并提供较长的读取长度。然而，迄今为止，纳米孔测序面临着许多技术挑战。杂交辅助纳米孔测序（HANS）方法克服了这些障碍。HANS利用探针库来检测目标DNA中的子序列，如杂交测序（SBH）。然而，HANS与SBH的不同之处在于，它也提取了位置信息，从而完全规避了SBH的局限性，使基因组长度测序成为可能。HANS平台将能够以低于1000美元的价格对人类基因组进行测序。此外，它还承诺能快速准确地做到这一点。我们的具体目的如下：1)合成和测试寡核苷酸标签的能力，以增强纳米孔检测寡核苷酸在目标DNA上的存在和确定寡核苷酸位置的能力。2)确定序列重建的最优算法方法，并对测序平台的性能特征进行估值。

DESCRIPTION (provided by applicant): The rationales for the development of technology that will enable extremely cheap, high speed sequencing are well established. Chief among these is the enablement of personalized medicine. There are currently in development several technologies that promise to markedly decrease the cost of sequencing a human genome. It is unclear, however, that any of these will be able to do so drastically enough to allow whole genome sequencing to become a routine clinical tool. Additionally, those technologies which are most promising on this cost parameter look as if they will face difficulties with respect to performance characteristics such as read length. One technology that promises to be cheap and fast and to provide long read lengths is nanopore-based sequencing. To date, however, nanopore sequencing has faced a number of technical challenges. The method of Hybridization-Assisted Nanopore Sequencing (HANS) overcomes these hurdles. HANS utilizes libraries of probes to detect subsequences in the target DNA as in sequencing by hybridization (SBH). HANS differs from SBH, however, in that positional information is also extracted thus completely circumventing, the limitations of SBH and making genome length sequencing feasible. The HANS platform will be capable of sequencing a human genome for substantially less than $1000. Additionally, it promises to do so quickly and accurately. Our specific aims are as follows: 1) Synthesize and test oligonucleotide tags for their ability to enhance the nanopore's capacity to detect the presence and determine the positions of the oligonucleotides on the target DNA. 2) Determine the optimal algorithmic approach for sequence reconstruction and estimate values for the performance characteristics of the sequencing platform.