被NIH称为革命性的下一代基因测序
转载请注明来自丁香园
Field: Multidisciplinary
Article Title: Accurate multiplex polony sequencing of an evolved bacterial genome
Authors: Shendure, J;Porreca, GJ;Reppas, NB;Lin, XX;McCutcheon, JP;Rosenbaum, AM;Wang, MD;Zhang, K;Mitra, RD;Church, GM
Journal: SCIENCE
Volume: 309
Issue: 5741
Page: 1728-1732
Q:Why do you think your paper is highly cited?
A：Yesterday’s genome projects have created an appetite for sequencing tomorrow, not easily handled by todays technology. This paper is an example of what NIH calls "revolutionary, next-generation sequencing." In addition, this paper is one of the first to show the power of lab evolution梒oupled to high-accuracy, whole-genome analysis.
Q: Does it describe a new discovery, methodology, or synthesis of knowledge?
A：This is about a method (or a few), but also about an attitude (of long-term technology commitment). The basic concepts of molecular multiplexing using "tags" for "genomic sequencing" has been around since our 1984 PNAS paper and the idea of molecular clones since the 1970s, but making a cell-free, electrophoresis-free, cloning and sequencing method that works well took a lot of new ideas.
Q: Could you summarize the significance of your paper in layman's terms?
A：Low-cost, high-accuracy genomes are arriving. We don’s have to wait for giant companies to create new technology or for giant centers to use it.
Q: How did you become involved in this research, and were any problems encountered along the way?
A：Start with an obsession with RNA & DNA structure in college and with getting everyone (who wants it) their own genome, add 30 years of trial-and-error, and a great community of like-minded folks and we are almost there.
Q: Are there any social or political implications for your research?
A：This enables the sequel to the Human Genome Project (HGP), the Personal Genome Project (PGP). Since DNA is increasingly "identifying," it may be wise to consent human subjects with clearer statements of the likely openness of their genomic data.
View a graphic of "Polony Sequencing Equipment".
George M. Church
Professor of Genetics, Harvard Medical School
MIT Health Sciences & Technology
Director of the Lipper Center for Computational Genetics
MIT-Harvard DOE Genomes to Life Center
NIH Center for Excellence in Genomic Science
Boston, MA, USA
丁香园网友[J_Lio]：
评论和导读：
这是篇05年9月发表在science上的关于DNA测序方法的文章。这种方法的诱人之处在于该方法非常便宜，并且精确性高，此外其所使用的试剂以及仪器都已经公开。因此可以预见，这种测序方法将会大有市场，按文中的原话来说，这种技术将会“commonly available”。
在开始翻译对话之前，让首先看看science上的原文：
原文连接：http://www.sciencemag.org/cgi/reprint/309/5741/1728.pdf
原文：
Accurate multiplex polony sequencing of an evolved bacterial genome
We describe a DNA sequencing technology in which a commonly available, inexpensive epifluorescence microscope is converted to rapid nonelectrophoretic DNA sequencing automation.We apply this technology to resequence an evolved
strain of Escherichia coli at less than one error per million consensus bases. A cell-free, mate-paired library provided single DNA molecules that were amplified in parallel to 1-micrometer beads by emulsion polymerase chain reaction. Millions of beads were immobilized in a polyacrylamide gel and subjected to automated cycles of sequencing by ligation and four-color imaging. Cost per base was roughly one-ninth as much as that of conventional sequencing. Our
protocols were implemented with off-the-shelf instrumentation and reagents.
翻译：
对纯化后的大肠杆菌菌株进行多元聚合扩增的高精度测序
在这篇文章中，研究人员开发了一种DNA测序技术，由于其比较便宜而且精确性很好，所以可以预见这种技术将会被普遍使用。此技术中主要使用了比较便宜的荧光显微镜，实现了非电泳的快速DNA自动测序。用这个技术对纯化后的大肠杆菌菌株再测序，结果错误率小于百万分之一。这种技术只需要双链DNA库，而无需细胞，测序的具体过程如下：首先扩增库中单条双链DNA分子，于是每条链用乳胶中聚合酶作用的方法被连接在一个微珠上，随后得到大量的微珠被固定到聚丙烯酰胺胶中，最后通过四色成像和分子间作用的方法进行自动测序。这种方法的花费仅仅是正常测序方法的九分之一。试验中的全部流程全部使用已经公开的仪器和试剂。
FBC 翻译：
问：您的这篇文章引用率很高，对此您有何看法？
答：我们之前已经完成的基因组计划激励着我们以后将要进行更深入的测序工作。而用今天的技术手段处理将要做的工作还十分棘手。我们这篇文章就提供了此方向的一个解决方法，同时这项研究被NIH（美国健康研究院）誉为“下一代测序手段的革命”。而且，使用实验进化的方法对高度精确的全基因组进行分析还是一个非常新的方向，这篇文章就是这个方向中重要的一篇。
问：可以称这篇文章中的内容为一种新的发现、方法、或者知识的综合吗？
答：我们这个研究不仅仅是涉及一种或一些方法，而且是一长期进行的技术研究。用标签（tags）作为衡量分子多样性的进本概念，进而进行基因组测序（genomic sequencing）的方法，始于我们1984年发表在PNAS上面的文章，这种方法还基于1970年后发展起来的分析克隆的方法。我们不通过细胞和电泳的手段进行克隆和测序的方法实际上涉及到了好多新方法。
问：请您简要的解释一下您在Layman的实验室中发表的这篇文章有什么样的重要性？
答：我们开发了一种真正的低花费，高精确性的全基因组测序手段。我们再也不必等待大公司开发新技术或者等待大型的实验中心使用这些技术（之后才能进行基因组测序实验了）。
问：您为什么要进行这方面的研究，在您研究过程中遇到过什么样的障碍和挫折？
答：在大学期间我就对RNA和DNA的结构比较着迷，而且有许多人希望得到他们自己的基因组序列，之后经历了30年的实验和失败，还有许许多多和我有着同样想法的人们一直在一起研究，这些都促使我从事这项研究。
问：您从事的这项研究将会给社会或者政府带来什么样的影响？
答：这项研究使得人类基因组计划（HGP）以及个人基因组计划（PGP）成为可能。随着越来越多的DNA序列被确定，我们就可以用已经公布的基因组数据来更深入地研究人类科学。
编辑：西门吹血
Article Title: Accurate multiplex polony sequencing of an evolved bacterial genome
Authors: Shendure, J;Porreca, GJ;Reppas, NB;Lin, XX;McCutcheon, JP;Rosenbaum, AM;Wang, MD;Zhang, K;Mitra, RD;Church, GM
Journal: SCIENCE
Volume: 309
Issue: 5741
Page: 1728-1732
Q:Why do you think your paper is highly cited?
A：Yesterday’s genome projects have created an appetite for sequencing tomorrow, not easily handled by todays technology. This paper is an example of what NIH calls "revolutionary, next-generation sequencing." In addition, this paper is one of the first to show the power of lab evolution梒oupled to high-accuracy, whole-genome analysis.
Q: Does it describe a new discovery, methodology, or synthesis of knowledge?
A：This is about a method (or a few), but also about an attitude (of long-term technology commitment). The basic concepts of molecular multiplexing using "tags" for "genomic sequencing" has been around since our 1984 PNAS paper and the idea of molecular clones since the 1970s, but making a cell-free, electrophoresis-free, cloning and sequencing method that works well took a lot of new ideas.
Q: Could you summarize the significance of your paper in layman's terms?
A：Low-cost, high-accuracy genomes are arriving. We don’s have to wait for giant companies to create new technology or for giant centers to use it.
Q: How did you become involved in this research, and were any problems encountered along the way?
A：Start with an obsession with RNA & DNA structure in college and with getting everyone (who wants it) their own genome, add 30 years of trial-and-error, and a great community of like-minded folks and we are almost there.
Q: Are there any social or political implications for your research?
A：This enables the sequel to the Human Genome Project (HGP), the Personal Genome Project (PGP). Since DNA is increasingly "identifying," it may be wise to consent human subjects with clearer statements of the likely openness of their genomic data.
View a graphic of "Polony Sequencing Equipment".
George M. Church
Professor of Genetics, Harvard Medical School
MIT Health Sciences & Technology
Director of the Lipper Center for Computational Genetics
MIT-Harvard DOE Genomes to Life Center
NIH Center for Excellence in Genomic Science
Boston, MA, USA
丁香园网友[J_Lio]：
评论和导读：
这是篇05年9月发表在science上的关于DNA测序方法的文章。这种方法的诱人之处在于该方法非常便宜，并且精确性高，此外其所使用的试剂以及仪器都已经公开。因此可以预见，这种测序方法将会大有市场，按文中的原话来说，这种技术将会“commonly available”。
在开始翻译对话之前，让首先看看science上的原文：
原文连接：http://www.sciencemag.org/cgi/reprint/309/5741/1728.pdf
原文：
Accurate multiplex polony sequencing of an evolved bacterial genome
We describe a DNA sequencing technology in which a commonly available, inexpensive epifluorescence microscope is converted to rapid nonelectrophoretic DNA sequencing automation.We apply this technology to resequence an evolved
strain of Escherichia coli at less than one error per million consensus bases. A cell-free, mate-paired library provided single DNA molecules that were amplified in parallel to 1-micrometer beads by emulsion polymerase chain reaction. Millions of beads were immobilized in a polyacrylamide gel and subjected to automated cycles of sequencing by ligation and four-color imaging. Cost per base was roughly one-ninth as much as that of conventional sequencing. Our
protocols were implemented with off-the-shelf instrumentation and reagents.
翻译：
对纯化后的大肠杆菌菌株进行多元聚合扩增的高精度测序
在这篇文章中，研究人员开发了一种DNA测序技术，由于其比较便宜而且精确性很好，所以可以预见这种技术将会被普遍使用。此技术中主要使用了比较便宜的荧光显微镜，实现了非电泳的快速DNA自动测序。用这个技术对纯化后的大肠杆菌菌株再测序，结果错误率小于百万分之一。这种技术只需要双链DNA库，而无需细胞，测序的具体过程如下：首先扩增库中单条双链DNA分子，于是每条链用乳胶中聚合酶作用的方法被连接在一个微珠上，随后得到大量的微珠被固定到聚丙烯酰胺胶中，最后通过四色成像和分子间作用的方法进行自动测序。这种方法的花费仅仅是正常测序方法的九分之一。试验中的全部流程全部使用已经公开的仪器和试剂。
FBC 翻译：
问：您的这篇文章引用率很高，对此您有何看法？
答：我们之前已经完成的基因组计划激励着我们以后将要进行更深入的测序工作。而用今天的技术手段处理将要做的工作还十分棘手。我们这篇文章就提供了此方向的一个解决方法，同时这项研究被NIH（美国健康研究院）誉为“下一代测序手段的革命”。而且，使用实验进化的方法对高度精确的全基因组进行分析还是一个非常新的方向，这篇文章就是这个方向中重要的一篇。
问：可以称这篇文章中的内容为一种新的发现、方法、或者知识的综合吗？
答：我们这个研究不仅仅是涉及一种或一些方法，而且是一长期进行的技术研究。用标签（tags）作为衡量分子多样性的进本概念，进而进行基因组测序（genomic sequencing）的方法，始于我们1984年发表在PNAS上面的文章，这种方法还基于1970年后发展起来的分析克隆的方法。我们不通过细胞和电泳的手段进行克隆和测序的方法实际上涉及到了好多新方法。
问：请您简要的解释一下您在Layman的实验室中发表的这篇文章有什么样的重要性？
答：我们开发了一种真正的低花费，高精确性的全基因组测序手段。我们再也不必等待大公司开发新技术或者等待大型的实验中心使用这些技术（之后才能进行基因组测序实验了）。
问：您为什么要进行这方面的研究，在您研究过程中遇到过什么样的障碍和挫折？
答：在大学期间我就对RNA和DNA的结构比较着迷，而且有许多人希望得到他们自己的基因组序列，之后经历了30年的实验和失败，还有许许多多和我有着同样想法的人们一直在一起研究，这些都促使我从事这项研究。
问：您从事的这项研究将会给社会或者政府带来什么样的影响？
答：这项研究使得人类基因组计划（HGP）以及个人基因组计划（PGP）成为可能。随着越来越多的DNA序列被确定，我们就可以用已经公布的基因组数据来更深入地研究人类科学。
编辑：西门吹血
作者: George M. Church
| 以下网友留言只代表网友个人观点，不代表网站观点 | |||
Copyright 2000-2025 DXY.CN All Rights Reserved