太平洋生物科学序列技术综述

Mohammed Abde Aliy, Senbeta Bayeta, Worku Takale
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引用次数: 0

摘要

太平洋生物科学公司开发了一种平台,可以通过聚合一种酶在一段时间内对一个DNA分子进行测序。太平洋生物科技的单分子实时测序技术是目前应用最为广泛的第三代测序技术之一。PacBio单分子实时测序利用零模式波导的独创性,从无序自由浮动的核苷酸产生的稳定荧光背景中区分出最佳荧光信号。PacBio单分子实时测序不需要PCR扩增,浏览长度比下一代测序长100倍。它将只覆盖高gc和高重复切片,在定量低频突变时更准确。PacBio单分子实时测序的错误率相对较高,为10%-15%(这实际上是现有单分子测序技术的标准缺陷)。然而,与下一代测序相比,这些错误无意中是随机的。因此,多次测序将有效地纠正底部偏差。与第二代测序不同,PacBio测序可能是一种周期测序技术,不需要在浏览步骤之间进行间歇。这些选项将PacBio测序与第二代测序区分开来,因此它被归类为第三代测序。PacBio测序产生极长的读取,错误率高,产率低。短读数将比对/组装/检测细化到单核苷酸精度,而PacBio长读数提供可靠的比对、支架和基因组变异的近似检测。PacBio测序系统通过超长的测序长度(平均100万bp)和高一致性精度,可以提供非常高的遗传信息深度。为了测量和推广PacBio测序信息分析的现代生物信息学工具,需要一台好的浏览机。
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Pacific bioscience sequence technology: Review
Pacific Biosciences has developed a platform that may sequence one molecule of DNA in a period via the polymerization of that strand with one enzyme. Single-molecule real-time sequencing by Pacific BioSciences’ technology is one of the most widely utilized third-generation sequencing technologies. PacBio single-molecule real-time Sequencing uses the Zero-mode waveguide’s ingenuity to distinguish the best fluorescence signal from the stable fluorescent backgrounds generated by disorganized free-floating nucleotides. PacBio single-molecule real-time sequencing does not require PCR amplification, and the browse length is a hundred times longer than next-generation sequencing. It will only cover high-GC and high-repeat sections and is more accurate in quantifying low-frequency mutations. PacBio single-molecule real-time sequencing will have a relatively high error rate of 10%-15% (which is practically a standard flaw of existing single-molecule sequencing technology). In contrast to next-generation sequencing, however, the errors are unintentionally random. As a result, multiple sequencing will effectively rectify the bottom deviance. Unlike second-generation sequencing, PacBio sequencing may be a technique for period sequencing and doesn’t need an intermission between browse steps. These options distinguish PacBio sequencing from second-generation sequencing, therefore it’s classified because of the third-generation sequencing. PacBio sequencing produces extremely lengthy reads with a high error rate and low yield. Short reads refine alignments/assemblies/detections to single-nucleotide precision, whereas PacBio long reads provide reliable alignments, scaffolds, and approximate detections of genomic variations. Through extraordinarily long sequencing reads (average >10,000 bp) and high accord precision, the PacBio Sequencing System can provide a terribly high depth of genetic information. To measure and promote the event of modern bioinformatics tools for PacBio sequencing information analysis, a good browse machine is required.
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