Name: denaturing urea polyacrylamide gel electrophoresis (urea page)
Uploaded: 2009-10-29T07:00:00
Description: Watch this Scientific Journal Video about Denaturing Urea Polyacrylamide Gel Electrophoresis Urea PAGE at JoVE.com

这项工作是由新加坡的学术研究理事会（ARC）[授予数量90/07]支持。我们感谢支持球菌。

为这一实验过程的完整和详细的文字协议是在分子生物学中的电流的协议。 BioRad公司网站3上可以找到详细的一步一步的组装凝胶三明治和凝胶仪器的指示。 所需设备： 玻璃板（内层和外层） 10厘米的电池：10.1 × 7.3厘米（内板），10.1 × 8.2厘米（外盘），BioRad公司 20厘米的电池：20 x 20厘米（内板），20 × 22.3厘米（外盘），BioRad公司 0.5-1.5毫米凝胶...

<ol>
	<li>Maniatis, T., Jeffrey, A., van deSande, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chain+length+determination+of+small+double-+and+single-stranded+DNA+molecules+by+polyacrylamide+gel+electrophoresis.">Chain length determination of small double- and single-stranded DNA molecules by polyacrylamide gel electrophoresis.</a> Biochemistry. 14, 3787-3794 (1975).</li><li>Albright, L. M., Slatko, B. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Denaturing+polyacrylamide+gel+electrophoresis.">Denaturing polyacrylamide gel electrophoresis.</a> Curr Protoc Nucleic Acid Chem. , (2001).</li><li>. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> PROTEAN II xi Cell and PROTEAN II xi 2-D Cell Instruction Manual. , (2001).</li></ol>

<ol><li>带的清晰度取决于几个因素。夏普带装样量应尽可能小，即理想之间的1-5μL。然而，到15μL可装载仍然确保接受的解决办法。样品量少，更清晰的频段量结果。 </li><li>该乐队的质量也取决于凝胶的厚度。稀释剂的凝胶，如0.75毫米，大于1.5毫米厚的凝胶显示较好的效果。 </li><li>乐队的形状也可以在凝胶加载的影响，如果样品是不能进入凝胶的口袋里同样适用。包括10％的装载缓冲甘油有助于在凝胶装载和更容?...

denaturing urea polyacrylamide gel electrophoresis (urea page)

尿素页面或变性尿素聚丙烯酰胺凝胶电泳采用6-8 M尿素，变性中学的DNA或RNA的结构，是他们根据分子量聚丙烯酰胺凝胶矩阵分离。 2至500个碱基之间的片段，如单核苷酸长度的差异，可以分开使用此方法1。样品的迁移是依赖于所选择的丙烯酰胺浓度。一个比例更高的聚丙烯酰胺解决了低分子量的碎片。尿素和温度为45-55 ° C进行组合，在凝胶运行允许非结构化的DNA或RNA分子的分离。 在一般情况下，这种方法是必需的单链DNA或RNA片段，合成或标记的寡核苷酸酶裂解反应的产品，如分析或净化。 在这个视频文章中，我们展示了如何准备和运行变性尿素聚丙烯酰胺凝胶。包括技术秘诀，除了原有的协议1,2。

Watch this Scientific Journal Video about Denaturing Urea Polyacrylamide Gel Electrophoresis Urea PAGE at JoVE.com

Denaturing Urea Polyacrylamide Gel Electrophoresis Urea PAGE

变性尿素聚丙烯酰胺凝胶电泳用于分离单链DNA或RNA的500个核苷酸的限制。在热变性样品和非结构化的单链结合的尿素凝胶基质内迁移，根据其分子量。

变性尿素聚丙烯酰胺凝胶电泳（尿素页）

Urea PAGE or denaturing urea polyacrylamide gel electrophoresis employs 6-8 M urea, which denatures secondary DNA or RNA structures and is used for their ...

Research

JoVE Journal

Biology

Denaturing Urea Polyacrylamide Gel Electrophoresis (Urea PAGE)

Denaturing Gradient Gel Electrophoresis (DGGE)

Denaturing Gradient Gel Electrophoresis DGGE

变性梯度凝胶电泳（DGGE）

科研

生物学

Optimized Fibrin Gel Bead Assay for the Study of Angiogenesis

优化的纤维蛋白凝胶珠检测血管生成的研究

Angiogenesis is a complex multi-step process, where, in response to angiogenic stimuli, new vessels are created from the existing vasculature.  These ...

Assessing Neural Stem Cell Motility Using an Agarose Gel-based Microfluidic Device

使用琼脂糖凝胶为基础的微流体装置的评估神经干细胞的运动

While microfluidic technology is reaching a new level of maturity for macromolecular assays, cell-based assays are still at an infant stage1. This is ...

Screening for Amyloid Aggregation by Semi-Denaturing Detergent-Agarose Gel Electrophoresis

半变性洗涤剂琼脂糖凝胶电泳筛选淀粉样蛋白聚合

Amyloid aggregation is associated with numerous protein misfolding pathologies and underlies the infectious properties of prions, which are ...

Fast and Sensitive Colloidal Coomassie G-250 Staining for Proteins in Polyacrylamide Gels

快速，灵敏的胶体考马斯亮G - 250染色蛋白聚丙烯酰胺凝胶

Coomassie Brilliant Blue (CBB) is a dye commonly used for the visualization of proteins separated by SDS-PAGE, offering a simple staining procedure and ...

Blue Native Polyacrylamide Gel Electrophoresis (BN-PAGE) for Analysis of Multiprotein Complexes from Cellular Lysates

Blue Native Polyacrylamide Gel Electrophoresis BN-PAGE for Analysis of Multiprotein Complexes from Cellular Lysates

蓝原生聚丙烯酰胺凝胶电泳（BN - PAGE）分析，从细胞裂解液多蛋白复合物

Multiprotein complexes (MPCs) play a crucial role in cell signalling, since most proteins can be found in functional or regulatory complexes with other ...

Pouring and Running a Protein Gel by reusing Commercial Cassettes

浇注和运行的一种蛋白质凝胶重用商​​业磁带

The evaluation of proteins using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis is a common technique used by biochemistry ...

Metabolic Pathway Confirmation and Discovery Through 13C-labeling of Proteinogenic Amino Acids

Metabolic Pathway Confirmation and Discovery Through 13C-labeling of Proteinogenic Amino Acids

代谢途径的确认和发现通过 13 C - Proteinogenic氨基酸的标签

Microbes have complex metabolic pathways that can be investigated using biochemistry and functional genomics methods. One important technique to examine ...

Agarose Gel Electrophoresis for the Separation of DNA Fragments

琼脂糖凝胶电泳分离DNA片段

Agarose gel electrophoresis is the most effective way of separating DNA fragments of varying sizes ranging from 100 bp to 25 kb1. Agarose is isolated from ...

Preparation of DNA-crosslinked Polyacrylamide Hydrogels

DNA交联聚丙烯酰胺水凝胶的制备

Mechanobiology is an emerging scientific area that addresses the critical role of physical cues in directing cell morphology and function. For example, ...