关键差异 - 水平与垂直凝胶电泳
凝胶电泳是一种实验室技术,用于分离含有的混合物DNA, RNA, and otherproteins根据它们各自的电荷和分子大小。在此方法中需要分离的DNA,RNA或蛋白质通过包含小毛孔的凝胶运行。分子通过电场通过凝胶驱动。分子穿过凝胶的孔,运动速度与它们各自的长度成反比。因此,分子大小较低的分子将比分子量较高的分子快速移动。电场是由凝胶两端的电荷差产生的。一端包含正电荷,另一端包含负电荷。由于DNA和RNA分子被负电荷,因此它们将被吸引到凝胶的正带端。凝胶电泳可以是两种不同的方法:水平凝胶电泳和垂直凝胶电泳。在水平凝胶电泳中,凝胶存在于水平方向上,并将其浸没在凝胶盒本身内的连续运行缓冲液中。在垂直凝胶电泳中,缓冲系统是垂直定向的,不连续,顶部和底部都有两个腔室,阴极和阳极, respectively。This is the key difference between horizontal and vertical gel electrophoresis.
内容
1.Overview and Key Difference
2.What is Horizontal Gel Electrophoresis
3.What is Vertical Gel Electrophoresis
4.水平和垂直凝胶电泳之间的相似性
5.Side by Side Comparison – Horizontal vs Vertical Gel Electrophoresis in Tabular Form
6。概括
What is Horizontal Gel Electrophoresis?
Horizontal gel electrophoresis utilizes the basic theory for separation of DNA, RNA or protein molecules according to their respective molecular size and charge. In this technique, the gel is present in a horizontal orientation and is submerged in a buffer which is continuous.Agarose凝胶用于将凝胶盒分成两个隔间。凝胶盒的一端包含一个阳极,而另一端则包含阴极。当应用电流时,该技术中使用的缓冲区允许创建电荷梯度。当施加电荷时,凝胶会倾向于加热。缓冲液还可以用作冷却液,该冷却液将温度保持在最佳水平。运行缓冲液的再循环防止pH梯度的形成。由于凝胶系统的两个隔室与运行缓冲液连接,因此不连续的缓冲系统无法在水平凝胶电泳中使用。Acrylamide在凝胶电泳期间用于分离蛋白质混合物。
In horizontal gel electrophoresis, acrylamide cannot be utilized since the gel box is exposed to oxygen. Due to the presence of oxygen, polymerization of acrylamide is inhibited, and this interferes with the formation of the gel. Horizontal gel electrophoresis is an effortless method that is utilized in the separation of DNA and RNA.
什么是垂直凝胶电泳?
Vertical gel electrophoresis technique functions according to the primary theory of gel electrophoresis, but it is considered to be more complex than horizontal gel electrophoresis method. This technique utilizes a discontinuous buffer. A cathode is located in the top chamber, and the anode is located in the bottom chamber. The electrodes present in each compartment provide the required electric field. A thin layer of gel is poured between the two mounted glass plates. Therefore, the top portion of the gel is submerged in the top chamber, and the bottom portion of the gel is submerged in the chamber at the bottom. Once the current is applied, a small portion of the buffer moves to the bottom chamber from the top chamber through the gel. The current applied in this technique is of minute units.
In vertical gel electrophoresis, buffer only flows through the gel. This allows accurate control of gradient of voltage during the separation stage. Acrylamide gel can be utilized since the compartments are not exposed to atmospheric oxygen. Due to the smaller pore size of acrylamide gel, precise separation can be achieved with a higher resolution.
水平和垂直凝胶电泳之间有什么相似之处?
- 两种系统都根据凝胶电泳的基本理论发挥作用。
- 阳极和阴极用于在两个系统中提供所需的电场。
水平和垂直凝胶电泳有什么区别?
Horizontal vs Vertical Gel Electrophoresis |
|
Horizontal Gel Electrophoresis is a gel electrophoresis technique in which the gel is present in a horizontal orientation. | 垂直凝胶电泳是一种凝胶电泳技术,其中凝胶垂直定向。 |
Buffer | |
水平凝胶电泳由连续缓冲液组成。 | 在垂直凝胶电泳中,运行缓冲液是不连续的。 |
Use of Acrylamide | |
Acrylamide cannot be utilized for horizontal gel electrophoresis since the gel box is exposed to atmospheric oxygen. | 由于凝胶由于两个单独的腔室而不会暴露于大气中的氧气中,因此可以将丙烯酰胺用于垂直凝胶电泳。 |
功能 | |
水平凝胶电泳更常用于DNA和RNA混合物的分离,而不是蛋白质的分离。 | Vertical gel electrophoresis is used to separate mixtures of protein. |
概括– Horizontal vs Vertical Gel Electrophoresis
凝胶电泳是实验室技术,可广泛用于含有DNA,RNA和蛋白质分子的混合物分离。凝胶电泳有两种方法:水平和垂直凝胶电泳。在水平凝胶电泳中,运行缓冲液是连续的,而在垂直凝胶电泳中,它是不连续的。这是水平和垂直凝胶电泳之间的差异。两种系统都根据凝胶电泳的共同原理发挥作用。
Download PDF Version of Horizontal vs Vertical Gel Electrophoresis
You can download PDF version of this article and use it for offline purposes as per citation note. Please download PDF version here水平和垂直凝胶电泳之间的差异。
参考:
1.沃伦,乍得,et al。”垂直琼脂糖凝胶electrophoresis and electroblotting of high-Molecular-Weight proteins.” Electrophoresis, vol. 24, no. 11, 2003, pp. 1695–1702., doi:10.1002/elps.200305392.
2.“Horizontal and Vertical Gel Systems – The Horizontal Gel System.” National Diagnostics,Available here。2017年8月28日访问。
Image Courtesy:
1. Jeffrey M. Vinocur的“凝胶电泳设备” - 自己的作品(CC BY 2.5)viaCommons Wikimedia
2.“蛋白质的聚丙烯酰胺凝胶电泳”,由Bruxelles的Jean-Etienne Minh-Duy Poirrier,比利时 - 一维凝胶电泳中的蛋白质(CC BY-SA 2.0)viaCommons Wikimedia
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