An investigative graduate laboratory course for teaching modern DNA techniques.
An investigative graduate laboratory course for teaching modern DNA techniques.
复制标题
DOI：
10.1002/bmb.21048
复制
发表时间：
2017-07-08
期刊：
影响因子：
--
通讯作者：
Keller LC
中科院分区：
文献类型：
--
作者：
de Lencastre A;Thomas Torello A;Keller LC
这个研究生水平的DNA方法实验室课程旨在模拟基于发现的研究项目，并让学生参与传统的DNA分析方法和现代重组DNA克隆技术。在课程的第一部分，学生使用Gateway®克隆技术克隆他们选择的人类疾病基因的果蝇直系同源物。在课程的第二部分，学生通过逆转录PCR检查他们感兴趣的基因在人类细胞系中的表达，并学习如何分析定量逆转录PCR（qRT-PCR）实验的数据。Gateway®克隆系统的适应性非常适合学生设计和创建不同类型的表达构建体，以实现特定的实验目标（例如蛋白质纯化，细胞培养中的表达和亚细胞定位），并且所选择的基因可以与教师的研究兴趣和/或部门正在进行的研究相一致。学生评估表明，该课程培养了人们对研究的真正兴奋，并深入了解了所执行的技术及其背后的理论。我们的长期目标是将这个DNA方法实验室作为昆尼皮亚克大学分子和细胞生物学理学硕士课程的综合实验室序列的基础，学生在随后的实验室课程中使用他们在本课程中开发的试剂和概念，包括蛋白质方法和细胞培养实验室。
This graduate-level DNA methods laboratory course is designed to model a discovery-based research project and engages students in both traditional DNA analysis methods and modern recombinant DNA cloning techniques. In the first part of the course, students clone the Drosophila ortholog of a human disease gene of their choosing using Gateway® cloning. In the second part of the course, students examine the expression of their gene of interest in human cell lines by reverse transcription PCR and learn how to analyze data from quantitative reverse transcription PCR (qRT-PCR) experiments. The adaptability of the Gateway® cloning system is ideally suited for students to design and create different types of expression constructs to achieve a particular experimental goal (e.g. protein purification, expression in cell culture, and subcellular localization), and the genes chosen can be aligned to the research interests of the instructor and/or ongoing research in a department. Student evaluations indicate that the course fostered a genuine excitement for research and in depth knowledge of both the techniques performed and the theory behind them. Our long-term goal is to incorporate this DNA methods laboratory as the foundation for an integrated laboratory sequence for the Master of Science degree program in Molecular and Cellular Biology at Quinnipiac University, where students use the reagents and concepts they developed in this course in subsequent laboratory courses, including a protein methods and cell culture laboratory.
登录
查看更多内容
DOI：
10.1002/bmb.20873
发表时间：
2015-07
期刊：
Biochemistry and molecular biology education : a bimonthly publication of the International Union of Biochemistry and Molecular Biology
影响因子：
--
作者：
Gray C;Price CW;Lee CT;Dewald AH;Cline MA;McAnany CE;Columbus L;Mura C
通讯作者：
Mura C
影响因子：
3.7
作者：
Cox-Paulson, Elisabeth A.;Grana, Theresa M.;Batzli, Janet M.
通讯作者：
Batzli, Janet M.
影响因子：
1.4
作者：
Carroll, Christopher W.;Keller, Lani C.
通讯作者：
Keller, Lani C.
影响因子：
3.7
作者：
Miller, Joanna A.;Witherow, D. Scott;Carson, Susan
通讯作者：
Carson, Susan
DOI：
10.1187/cbe.04-09-0052
发表时间：
2005-01-01
期刊：
Cell biology education
影响因子：
--
作者：
Howard, David R;Miskowski, Jennifer A
通讯作者：
Miskowski, Jennifer A