Denaturing gel electrophoresis for sequencing.
Denaturing gel electrophoresis for sequencing.
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用于测序的变性凝胶电泳。
DOI：
--
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发表时间：
2001
影响因子：
--
通讯作者：
L. M. Albright
中科院分区：
文献类型：
--
作者：
B. Slatko;L. M. Albright
DNA序列测定的准确性很大程度上取决于变性聚丙烯酰胺凝胶中测序产物的分辨率。本单元提供了设置，电泳和处理这些凝胶的详细说明。一般来说，DNA测序所需的凝胶长40厘米，厚度均匀，含有4%至8%的丙烯酰胺和7 M尿素。该方案的修改增加了可从单个凝胶获得的可读序列信息的长度（即，形成具有楔形间隔物的凝胶以产生场梯度，或将缓冲液梯度、电解质梯度或丙烯酰胺阶梯梯度掺入凝胶中）。对基本方案的修改-在测序凝胶中包含甲酰胺-旨在克服凝胶电泳期间测序产物中二级结构引起的凝胶压缩。丙烯酰胺浓度和电泳条件的讨论包括在评注中。
The accuracy of DNA sequence determination depends largely upon resolution of the sequencing products in denaturing polyacrylamide gels. This unit provides a detailed description of the setup, electrophoresis, and processing of such gels. In general, the gels required for DNA sequencing are 40-cm long, of uniform thickness, and contain 4% to 8% acrylamide and 7 M urea. Modifications of this protocol increase the length of readable sequence information which can be obtained from a single gel (i.e., forming the gel with wedge-shaped spacers to create a field gradient, or incorporating a buffer gradient, an electrolyte gradient, or an acrylamide step gradient into the gel). A modification to the Basic Protocol--inclusion of formamide in the sequencing gel--is designed to overcome gel compressions arising from secondary structure in the sequencing products during gel electrophoresis. A discussion of acrylamide concentrations and electrophoresis conditions is included in the Commentary.
DOI：
10.1073/pnas.84.14.4767
发表时间：
1987-07-01
影响因子：
11.1
作者：
TABOR, S;RICHARDSON, CC
通讯作者：
RICHARDSON, CC
序列分析的4-6-8方法。
影响因子：
2.7
作者：
Isfort,RJ;Ihle,JN
通讯作者：
Ihle,JN