DNA labeling for improved synthetic long reads

项目摘要 长读测序有可能极大地简化测序，有助于加快科学家的 能够进行从头测序、单倍型分期和转录组分。该项目旨在开发一种 在下一代测序之前标记DNA的方法，该方法维护关于 在原始链中的片段，有助于序列数据的下游组装。为了标记DNA， 转座酶将携带含有条形码标签的特殊设计的转座子，并用于片段 DNA，在下一代测序之前。这个项目的第一个目标是构建所使用的转座体 来标记DNA。第二个目标是使用这些转座体来标记和测序一个模型DNA系统 使用下一代测序在不到1皮克的DNA上演示~50kb的读出长度。如果 成功，在这笔赠款中开发的测序方法将简化合成长读测序，使 高精确度和廉价的长阅读测序更容易为遗传学社区所接受。

Project Summary Long-read sequencing has the potential to greatly simplify sequencing, helping to accelerate scientists’ ability to perform de novo sequencing, haplotype phasing and transcriptomics. This project aims to develop a method to label DNA prior to next-generation sequencing, that maintains information about the proximity of fragments in the original strand, aiding in the downstream assembly of the sequencing data. To label the DNA, transposase will be loaded with specially-designed transposons containing barcode labels, and used to fragment the DNA, prior to next-generation sequencing. The first aim of this project is to construct the transposomes used to label the DNA. The second aim is to use these transposomes to tagment and sequence a model DNA system to demonstrate read lengths of ~50 kb using next-generation sequencing on less than a picogram of DNA. If successful, the sequencing approach developed in this grant will simplify synthetic long read sequencing, making high-accuracy and inexpensive long read sequencing more accessible to the genetics community.