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» 电泳

电泳

1970
  • Arne Tiselius
按大小分离 DNA 样品的实验室凝胶电泳仪。.

(图片仅供参考)

凝胶电泳是一种根据DNA、RNA和蛋白质等大分子的大小和电荷进行分离的技术。将电场施加到凝胶基质上,使带负电荷的分子(如DNA)向正极移动。较小的分子比较大的分子在凝胶孔隙中迁移得更快、更远。

Gel electrophoresis is a cornerstone technique in molecular biology for the separation and analysis of macromolecules, primarily DNA, RNA, and proteins. The principle behind the technique is the migration of charged molecules through a porous gel matrix under the influence of an electric field. Since nucleic acids (DNA and RNA) have a consistently negative charge due to their phosphate backbone, they will naturally migrate towards the positive electrode (anode) when a current is applied. The gel itself, typically made of agarose for larger molecules like genomic DNA fragments or polyacrylamide for smaller fragments and proteins (in a technique called PAGE), acts as a molecular sieve. The matrix is composed of a network of pores through which the molecules must travel. Smaller molecules navigate this maze more easily and quickly than larger molecules, which are impeded more by the gel matrix. Consequently, the molecules are separated based on their size, with the smallest traveling the farthest from the starting point (the well) in a given amount of time. To perform DNA electrophoresis, a sample is first loaded into wells at one end of the gel. A ‘DNA ladder,’ a mixture of DNA fragments of known sizes, is run in a separate lane to serve as a reference for estimating the size of the unknown fragments. After the electric current is applied for a period, the separated fragments are visualized. Because DNA is invisible to the naked eye, a fluorescent dye that intercalates with the DNA, such as ethidium bromide or SYBR Green, is used. When placed under UV light, the dye fluoresces, revealing the DNA as distinct bands, each band representing a collection of fragments of the same size.

UNESCO Nomenclature: 2401
- 生物化学

类型

物理过程

中断

基础

用法

广泛使用

前体

  • 迈克尔-法拉第的电解定律
  • 电的发现
  • Arne Tiselius 开发的移动边界电泳
  • 开发支持介质,如纸、淀粉,以及琼脂糖/聚丙烯酰胺凝胶
  • DNA及其负电荷的发现

应用程序

  • DNA指纹识别
  • 亲子鉴定
  • 诊断遗传疾病
  • 纯化用于克隆的 DNA 片段
  • 分析PCR产物
  • 蛋白质分析(SDS-PAGE)

专利:

NA

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相关术语:凝胶电泳、DNA分离、琼脂糖凝胶、聚丙烯酰胺、SDS-PAGE、DNA指纹图谱、分子量、限制性片段、核酸、蛋白质。

历史背景

电泳

1954
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1951
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1975
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1983

(如果日期未知或不相关,例如“流体力学”,则提供其显著出现的近似估计)

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