Light Kylie, Brooks Jordan, Choi Yun-Seok
School of Natural Sciences, Black Hills State University, Spearfish, South Dakota 57799, United States.
J Chem Educ. 2023 Jul 20;100(8):3138-3143. doi: 10.1021/acs.jchemed.3c00177. eCollection 2023 Aug 8.
Sanger sequencing, also known as dideoxy sequencing, is a widely used method for DNA sequencing, particularly for cloned plasmids and clinical samples. This technique requires a combination of essential biochemistry skills, such as a chain-termination reaction, gel electrophoresis, and fluorescence detection. Unfortunately, there is a lack of activities that replicate the Sanger sequencing process for students to learn and practice these skills. To address this issue, a manipulative-based Sanger sequencing activity was developed that incorporates colorful pop beads to demonstrate a chain-termination reaction, separation of products, and fluorescence detection. The beads represent deoxynucleotides and dideoxynucleotides, allowing for a visual representation of DNA fragment generation. This kinesthetic learning activity offers a high visual impact for students, aiding in their understanding of the Sanger sequencing process, and can also be used to illustrate polymerase chain reaction (PCR)-based techniques.
桑格测序法,也称为双脱氧测序法,是一种广泛应用于DNA测序的方法,尤其适用于克隆质粒和临床样本。该技术需要综合运用基本的生物化学技能,如链终止反应、凝胶电泳和荧光检测。遗憾的是,缺乏能让学生学习和练习这些技能的、模拟桑格测序过程的活动。为了解决这个问题,开发了一种基于操作的桑格测序活动,该活动采用彩色的珠子来演示链终止反应、产物分离和荧光检测。这些珠子代表脱氧核苷酸和双脱氧核苷酸,能够直观呈现DNA片段的生成。这种动觉学习活动对学生具有很高的视觉冲击力,有助于他们理解桑格测序过程,还可用于阐释基于聚合酶链反应(PCR)的技术。