Goodwin Sara, Wappel Robert, McCombie W Richard
Genome Center, Cold Spring Harbor Laboratory, Cold Spring Harbor, New York.
Curr Protoc Hum Genet. 2017 Jul 11;94:18.11.1-18.11.14. doi: 10.1002/cphg.39.
Today's short-read sequencing instruments can generate read lengths between 50 bp and 700 bp depending on the specific instrument. These high-throughput sequencing approaches have revolutionized genomic science, allowing hundreds of thousands of full genomes to be sequenced, and have become indispensable tools for many researchers. With greater insight has come the revelation that many genomes are much more complicated than originally thought and include many rearrangements and copy-number variations. Unfortunately, short-read sequencing technologies are not well suited for identifying many of these types of events. Long-read sequencing technologies can read contiguous fragments of DNA in excess of 10 kb and are much better suited for detecting large structural events. The newest long-read sequencing instrument is the MinION device from Oxford Nanopore. The rapid sequencing speed and low upfront instrument cost are features drawing interest in this device from the genomics community. This unit provides a representative protocol for carrying out human genome sequencing on the Oxford Nanopore MinION. © 2017 by John Wiley & Sons, Inc.
如今的短读长测序仪器根据具体仪器不同，能产生50碱基对至700碱基对之间的读长。这些高通量测序方法彻底改变了基因组科学，使得数十万完整基因组得以测序，并且已成为许多研究人员不可或缺的工具。随着认识的深入，人们发现许多基因组比最初认为的要复杂得多，包含许多重排和拷贝数变异。不幸的是，短读长测序技术不太适合识别许多这类事件。长读长测序技术能够读取超过10 kb的连续DNA片段，更适合检测大型结构事件。最新的长读长测序仪器是牛津纳米孔公司的MinION设备。快速的测序速度和较低的仪器前期成本是该设备吸引基因组学界关注的特点。本单元提供了一个在牛津纳米孔MinION上进行人类基因组测序的代表性方案。© 2017约翰威立国际出版公司
