关键区别 - 微阵列与下一代测序
DNA测序processes are used widely in the fields of生物技术, virology, medical diagnosis and forensic sciences. It is a process which determines the exact order of the核苷酸,腺嘌呤、鸟嘌呤,胸骨和cytosinepresent within a DNA molecule. DNA sequencing procedures have become an accelerant for miraculous discoveries in medical and biological research. These sequencing methods have evolved up to sequencing a complete genome of individual organisms including humans and other living species. Microarrays and Next Generation Sequencing are modern DNA sequencing procedures.微阵列技术专门基于hybridization其中包含一组已知目标。下一代测序基于合成(利用DNA聚合酶掺入核苷酸),并具有独立于先前选择靶标的整个基因组对整个基因组进行测序的能力。这是微阵列和下一代测序之间的关键区别。
内容
1.Overview and Key Difference
2.What is Microarray
3.什么是下一代测序
4.微阵列和下一代测序之间的相似之处
5。并排比较 - 以表格形式进行微阵列与下一代测序
6。概括
什么是微阵列?
DNA微阵列被用作实验室工具,以同时识别数千种不同的基因表达。它是一个固体表面,即显微镜载玻片,其中包含在其上印有微观DNA斑点的集合。每个印刷的点都包含已知的基因序列或一个基因。这些已知probesprinted on the slide serve as probes in order to detectgene expression。This is known as atranscriptome。两个DNA链之间的杂交是主要的原理微阵列。它是核酸序列的互补碱基配对与形成氢键。
Initially,mRNAmolecules are collected from the experimental sample and reference sample obtained from a healthy individual. Experimental samples are obtained from diseased individuals; for example, an individual suffering from cancer. Once obtained, both mRNA samples are converted tocDNA(complementary DNA). Next, each sample is labeled using a fluorescent probe. The fluorescent probes are of different colors to distinguish the sample cDNA from the reference cDNA. In order to initiate the binding of the cDNA molecules to the microarray slide, the two samples are mixed together. Hybridization is the process by which the cDNA molecules get attached to the DNA probes on the microarray slide. Once hybridization is completed, a series of reactions take place in order to identify and measure the expression of each gene with the appearance of different colors according to the amount of the gene expressed. The results from microarray are utilized in the creation of a gene expression profile which can be used to identify different disease conditions.
什么是下一代测序?
Next Generation Sequencing (NGS) is an advanced method of genetic sequencing. Its principle is similar to that ofSanger Sequencing,取决于毛细管电泳。在NGS中,基因组链被碎裂并连接到模板链。每个链的底座在连接过程中由发射信号识别。在Sanger测序方法中,涉及三个单独的步骤,测序,分离和检测。由于这些单独的步骤,样品制备的自动化在吞吐量上受到限制。在NGS中,该技术是使用基于阵列的测序与Sanger测序过程的组合进行开发的,该步骤可能会导致数百万个反应序列同时进行。这导致高速和吞吐量以低成本。
NGS is composed of three steps; library preparation (creation of libraries with the use of random fragmentation of DNA), amplification (amplification of the library using clonal amplification andpcr)和测序。使用Sanger测序程序在极长的持续时间内进行的基因组测序过程可以在几个小时内使用NGS完成。
What is the Similarity Between Microarray and Next Generation Sequencing?
- Both Microarray and Next Generation Sequencing are developed using array based sequencing.
微阵列和下一代测序之间有什么区别?
Microarray vs Next Generation Sequencing |
|
微阵列是附着在固体表面上的微观DNA斑点的集合,该斑点用于同时测量大量基因的表达水平。 | NGS(下一代测序)是一种基于非距离的高通量DNA测序技术,可促进数百万或数十亿或数十亿的DNA链,以平行测序。 |
与抗原的相互作用 | |
微阵列基于由一组已知靶标组成的杂交。 | NGS基于合成,该合成利用DNA聚合酶掺入核苷酸,并且独立于先前选择的靶标。 |
摘要 - 微阵列与下一代测序
In the context of research, DNA sequencing has become an important accelerant. It is widely used in biotechnology, medical diagnosis, and forensic studies. It has evolved and developed into more efficient and rapid sequencing procedures. Microarrays and NGS are two advanced DNA sequencing techniques present. Both are developed using array based sequencing. Microarray technique relies on hybridization whilst NGS is based on synthesis, which utilizes DNA polymerase to incorporate nucleotides. This is the main difference between Microarray and Next Generation Sequencing.
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参考:
1.Behjati, Sam, and Patrick S Tarpey. “What is next generation sequencing?”Archives of Disease in Childhood. Education and Practice Edition, BMJ Publishing Group, Dec. 2013,Available here.Accessed 23 Aug. 2017
2.Bumgarner, Roger. “Overview of dna microarrays.” Current Protocols in Molecular Biology, John Wiley and Sons Inc., 9 Feb. 2016,Available here。2017年8月23日访问。
3.“ DNA微阵列技术”。国家人类基因组研究所(NHGRI),Available here.2017年8月23日访问。
Image Courtesy:
1. “DNA microarray” By Guillaume Paumier (user:guillom) – Own work(CC BY-SA 3.0)viaCommons Wikimedia
2. “下一代测序的发展” By Nederbragt, Lex (2012) –(CC BY 3.0)viaCommons Wikimedia
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