PacBio Sequencing and Its Applications.
PacBio Sequencing and Its Applications.
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DOI：
10.1016/j.gpb.2015.08.002
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发表时间：
2015-10
期刊：
影响因子：
--
通讯作者：
Au KF
中科院分区：
文献类型：
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作者：
Rhoads A;Au KF
Pacific BioSciences开发的单分子实时测序技术比第二代测序技术（SGS）提供了更长的读取长度，使其非常适合基因组、转录组和表观遗传学研究中尚未解决的问题。使用PacBio测序的高度连续的从头组装可以填补现有参考组装的空白，并表征个人基因组的结构变异（SV）。通过更长的读取，我们可以通过扩展的重复区域进行测序并检测突变，其中许多突变与疾病有关。此外，由于PacBio转录组测序能够对全长转录本或长度较大的片段进行测序，因此有利于鉴定基因同工型，并有助于可靠地发现新基因和注释基因的新同工型。此外，PacBio的测序技术为直接检测碱基修饰（如甲基化）提供了有用的信息。除了单独使用PacBio测序外，许多混合测序策略已经开发出来，以使用更精确的短读取与PacBio长读取相结合。一般来说，混合测序策略比单独使用PacBio测序更经济、更可扩展，特别是对于小型实验室。PacBio测序技术的出现提供了许多仅通过SGS无法获得的信息。
Single-molecule, real-time sequencing developed by Pacific BioSciences offers longer read lengths than the second-generation sequencing (SGS) technologies, making it well-suited for unsolved problems in genome, transcriptome, and epigenetics research. The highly-contiguous de novo assemblies using PacBio sequencing can close gaps in current reference assemblies and characterize structural variation (SV) in personal genomes. With longer reads, we can sequence through extended repetitive regions and detect mutations, many of which are associated with diseases. Moreover, PacBio transcriptome sequencing is advantageous for the identification of gene isoforms and facilitates reliable discoveries of novel genes and novel isoforms of annotated genes, due to its ability to sequence full-length transcripts or fragments with significant lengths. Additionally, PacBio’s sequencing technique provides information that is useful for the direct detection of base modifications, such as methylation. In addition to using PacBio sequencing alone, many hybrid sequencing strategies have been developed to make use of more accurate short reads in conjunction with PacBio long reads. In general, hybrid sequencing strategies are more affordable and scalable especially for small-size laboratories than using PacBio Sequencing alone. The advent of PacBio sequencing has made available much information that could not be obtained via SGS alone.