Man Y G, Kuhls E A, Bratthauer G L, Moinfar F, Tavassoli F A
Department of Gynecologic and Breast Pathology, Armed Forces Institute of Pathology, American Registry of Pathology, Washington, DC 20306-6000, USA.
Electrophoresis. 2001 Jun;22(10):1915-9. doi: 10.1002/1522-2683(200106)22:10<1915::AID-ELPS1915>3.0.CO;2-9.
Attempting to assess whether a decrease of the electrophoresis temperature could prevent or reduce the extent of gel well deformations, and whether the utilization of native polyacrylamide gels (without urea) could speed up the separation of polymerase chain reaction (PCR)-amplified products with an automated 377 DNA sequencer, denatured PCR products were subjected to electrophoresis in 6% native gels under 45 degrees C. Results show that a decrease of the electrophoresis temperature from 51 degrees C (recommended by the User's Manual) to 45 degrees C substantially facilitates the preservation of gel wells, and that all PCR products tested migrate significantly faster in native than in denatured (with urea) gels of the same concentration. The combination of a 6% native gel and a lower (45 degrees C) electrophoresis temperature permits multiple uses of a given gel with consistent results, consequently reducing the electrophoresis time and reagent costs.
为了评估降低电泳温度是否可以防止或减少凝胶孔变形的程度，以及使用天然聚丙烯酰胺凝胶（不含尿素）是否可以通过自动377 DNA测序仪加快聚合酶链反应（PCR）扩增产物的分离，将变性的PCR产物在45℃下于6%的天然凝胶中进行电泳。结果表明，将电泳温度从51℃（用户手册推荐）降至45℃可显著促进凝胶孔的保存，并且所有测试的PCR产物在天然凝胶中的迁移速度明显快于相同浓度的变性（含尿素）凝胶。6%天然凝胶和较低（45℃）电泳温度的组合允许对给定凝胶进行多次使用且结果一致，从而减少了电泳时间和试剂成本。
