Fredlake Christopher P, Hert Daniel G, Mardis Elaine R, Barron Annelise E
Department of Chemical and Biological Engineering, Northwestern University, Evanston, IL, USA.
Electrophoresis. 2006 Oct;27(19):3689-702. doi: 10.1002/elps.200600408.
Although a finished human genome reference sequence is now available, the ability to sequence large, complex genomes remains critically important for researchers in the biological sciences, and in particular, continued human genomic sequence determination will ultimately help to realize the promise of medical care tailored to an individual's unique genetic identity. Many new technologies are being developed to decrease the costs and to dramatically increase the data acquisition rate of such sequencing projects. These new sequencing approaches include Sanger reaction-based technologies that have electrophoresis as the final separation step as well as those that use completely novel, nonelectrophoretic methods to generate sequence data. In this review, we discuss the various advances in sequencing technologies and evaluate the current limitations of novel methods that currently preclude their complete acceptance in large-scale sequencing projects. Our primary goal is to analyze and predict the continuing role of electrophoresis in large-scale DNA sequencing, both in the near and longer term.
尽管现在已有完整的人类基因组参考序列,但对生物科学领域的研究人员来说,对大型复杂基因组进行测序的能力仍然至关重要。特别是,持续进行人类基因组序列测定最终将有助于实现根据个体独特的基因特征量身定制医疗保健的愿景。目前正在开发许多新技术,以降低此类测序项目的成本并大幅提高数据采集率。这些新的测序方法包括以桑格反应为基础、以电泳作为最终分离步骤的技术,以及那些使用全新的非电泳方法来生成序列数据的技术。在这篇综述中,我们讨论了测序技术的各种进展,并评估了目前新方法存在的局限性,这些局限性使得它们目前无法在大规模测序项目中被完全接受。我们的主要目标是分析和预测电泳在近期和更长期的大规模DNA测序中持续发挥的作用。