The "Frankenplasmid" Lab AN INVESTIGATIVE EXERCISE FOR TEACHING RECOMBINANT DNA METHODS
The "Frankenplasmid" Lab AN INVESTIGATIVE EXERCISE FOR TEACHING RECOMBINANT DNA METHODS
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DOI：
10.1002/bmb.20531
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发表时间：
2011-09-01
影响因子：
1.4
通讯作者：
Wilder, Jason A.
中科院分区：
文献类型：
--
作者：
Dean, Derek M.;Wilder, Jason A.
我们描述了一个调查实验室模块，旨在让大学本科生在3周内重组DNA方法的实践和理论经验。在推导出两种不同质粒的限制性内切酶图谱后，学生在同一反应中将质粒连接在一起，转化E。用这种连接的DNA混合物感染大肠杆菌，并将细胞接种在特异性选择杂交质粒的培养基上。作业的主要目的是让学生利用其转化体的LacZ表型、PCR和限制性内切酶图谱来推导一个杂交“弗兰肯质粒”的基因图谱。我们的方案产生了许多可能的结果，这意味着学生们正在绘制真正未知的质粒。这项任务的开放性导致了一个有效的模块，该模块教授重组DNA程序，同时让学生参与其调查方法，增加复杂性和谜题般的质量。此外，活动的模块化设计使其能够适应更有限的时间表，入门课程或更高级的课程。
We describe an investigative laboratory module designed to give college undergraduates strong practical and theoretical experience with recombinant DNA methods within 3 weeks. After deducing restriction enzyme maps for two different plasmids, students ligate the plasmids together in the same reaction, transform E. coli with this mixture of ligated DNA, and plate the cells on media that specifically select for hybrid plasmids. The main goal of the assignment is for students to deduce the gene map of one hybrid "Frankenplasmid" using the LacZ phenotype of its transformants, PCR, and restriction mapping. Our protocol results in a number of possible outcomes, meaning that students are mapping truly unknown plasmids. The open-ended nature of this assignment results in an effective module that teaches recombinant DNA procedures while engaging students with its investigative approach, increasing complexity, and puzzle-like quality. Moreover, the modular design of the activity allows it to be adapted to a more limited schedule, introductory courses, or more advanced courses.