Bradford William D, Cahoon Laty, Freel Sara R, Hoopes Laura L Mays, Eckdahl Todd T
Missouri Western State College, Saint Joseph, MO 64507, USA.
Cell Biol Educ. 2005 Summer;4(2):157-68. doi: 10.1187/cbe.04-09-0051.
In order to engage their students in a core methodology of the new genomics era, an ever-increasing number of faculty at primarily undergraduate institutions are gaining access to microarray technology. Their students are conducting successful microarray experiments designed to address a variety of interesting questions. A next step in these teaching and research laboratory projects is often validation of the microarray data for individual selected genes. In the research community, this usually involves the use of real-time polymerase chain reaction (PCR), a technology that requires instrumentation and reagents that are prohibitively expensive for most undergraduate institutions. The results of a survey of faculty teaching undergraduates in classroom and research settings indicate a clear need for an alternative approach. We sought to develop an inexpensive and student-friendly gel electrophoresis-based PCR method for quantifying messenger RNA (mRNA) levels using undergraduate researchers as models for students in teaching and research laboratories. We compared the results for three selected genes measured by microarray analysis, real-time PCR, and the gel electrophoresis-based method. The data support the use of the gel electrophoresis-based method as an inexpensive, convenient, yet reliable alternative for quantifying mRNA levels in undergraduate laboratories.
为了让学生参与新基因组学时代的核心方法，越来越多主要为本科院校的教师能够使用微阵列技术。他们的学生正在成功开展旨在解决各种有趣问题的微阵列实验。这些教学和研究实验室项目的下一步通常是对个别选定基因的微阵列数据进行验证。在研究领域，这通常涉及使用实时聚合酶链反应（PCR），这项技术所需的仪器和试剂对大多数本科院校来说过于昂贵。一项针对在课堂和研究环境中教授本科生的教师的调查结果表明，显然需要一种替代方法。我们试图开发一种基于凝胶电泳的廉价且对学生友好的PCR方法，以使用本科研究人员作为教学和研究实验室中学生的模型来定量信使核糖核酸（mRNA）水平。我们比较了通过微阵列分析、实时PCR和基于凝胶电泳的方法测量的三个选定基因的结果。数据支持将基于凝胶电泳的方法作为本科实验室中定量mRNA水平的一种廉价、便捷且可靠的替代方法。
