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1. 发明申请

US20020128415A1 Polyacrylamide medium for the electrophoretic separation of biomolecules 失效
标题翻译：用于电泳分离生物分子的聚丙烯酰胺培养基
公开(公告)号：US20020128415A1
公开(公告)日：2002-09-12
申请号：US10094397
申请日：2002-03-07
申请人： The Regents of the University of California
发明人： Ramakrishna S. Madabhushi , Stuart A. Gammon
IPC分类号： C08F120/54
CPC分类号： C08F120/54 , G01N27/44747
摘要： A polyacryalmide medium for the electrophoretic separation of biomolecules. The polyacryalmide medium comprises high molecular weight polyacrylamides (PAAm) having a viscosity average molecular weight (Mv) of about 675-725 kDa were synthesized by conventional red-ox polymerization technique. Using this separation medium, capillary electrophoresis of BigDye DNA sequencing standard was performed. A single base resolution of null725 bases was achieved in null60 minute in a non-covalently coated capillary of 50 nullm i.d., 40 cm effective length, and a filed of 160 V/cm at 40null C. The resolution achieved with this formulation to separate DNA under identical conditions is much superior (725 bases vs. 625 bases) and faster (60 min. vs. 75 min.) to the commercially available PAAm, such as supplied by Amersham. The formulation method employed here to synthesize PAAm is straight-forward, simple and does not require cumbersome methods such as emulsion polymerizaiton in order to achieve very high molecular weights. Also, the formulation here does not require separation of PAAm from the reaction mixture prior to reconstituting the polymer to a final concentration. Furthermore, the formulation here is prepared from a single average mol. wt. PAAm as opposed to the mixture of two different average mo. wt. PAAm previously required to achieve high resolution.
摘要翻译：用于电泳分离生物分子的聚丙烯酰胺培养基。聚丙烯酰胺介质包含通过常规红 - 氧聚合技术合成的具有约675-725kDa粘度平均分子量（Mv）的高分子量聚丙烯酰胺（PAAm）。使用该分离介质，进行BigDye DNA测序标准的毛细管电泳。在50mil，40cm有效长度的非共价涂布的毛细管中，在40℃下，160V / cm的非共价涂层毛细管在〜60分钟内实现约725个碱基的单碱基分辨率。用该配方实现的分辨率在相同条件下分离DNA比（例如由Amersham提供的）更优越（725碱基对625碱基）和更快（60分钟，相对于75分钟）的市售PAAm。这里用于合成PAAm的制剂方法是直接的，简单的，并且不需要繁琐的方法，例如乳液聚合，以获得非常高的分子量。此外，这里的配方不需要在将聚合物重构成最终浓度之前将PAAm与反应混合物分离。此外，这里的配方由单一平均摩尔数制备。重量 PAAm与两种不同平均水平的混合物相反。重量 PAAm以前需要达到高分辨率。

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