化学生物学实验“DNA片段扩增和琼脂糖凝胶电泳检测”的改进研究  

Research on Improvement of Chemical Biology Experiment"Amplification of DNA Fragments and Agarose Gel Electrophoresis Detection

作　　者：包婷[1] 熊成义 文为[1] 张修华[1] 何汉平[1] 王升富[1] Bao Ting;Xiong Cheng-yi;Wen Wei;Zhang Xiu-hua;He Han-ping;Wang Sheng-fu(College of Chemistry and Chemical Engineering,Hubei University,Wuhan 430062,China)

机构地区：[1]湖北大学化学化工学院,湖北武汉430062

出　　处：《化学传感器》2022年第3期52-55,共4页Chemical Sensors

基　　金：湖北大学2022年度教学改革研究项目“基于SPOC线上线下混合式教学的地方性高校化学类综合实验的重塑和优化(编号2022008)

摘　　要：DNA片段扩增和琼脂糖凝胶电泳检测是化学生物学的一个重要专业综合实验,旨在培养学生的综合实验技能并拓展科学思维。通常情况下,该实验利用聚合酶链式反应(PCR)进行DNA片段的扩增,但PCR仪器和试剂较为昂贵,大幅增加了实验成本。此外,PCR操作的技术性较强,操作繁琐并涉及到多种微量试剂的移取,实验误差较大导致难以得到令人满意的实验结果。针对上述问题,该文提出利用杂交链式反应(HCR)代替PCR法进行DNA片段的扩增,该方法为无酶等温核酸扩增技术,简单、温和且不需要昂贵的设备。此外,通过指导学生进行DNA序列的设计,可以加深对HCR原理、碱基互补配对原则的理解和应用。基于HCR的DNA片段扩增避免了昂贵仪器和试剂的使用,降低了实验成本,简化了实验操作,提高了实验的成功率。Amplification of DNA fragments and agarose gel electrophoresis detection is an important professional comprehensive experiment in chemical biology,which aims to cultivate the comprehensive experimental skills and expand scientific cogitation of students.Usually,this experiment utilizes polymerase chain reaction(PCR)to amplify DNA fragments,however,the instrument and reagents of PCR are relatively expensive,which greatly increases the experimental cost.In addition,the PCR operation is relatively highly technical,the operation is cumbersome and involved in the pipetting of a variety of trace reagents.Therefore,the large experimental error makes it difficult to obtain satisfactory experimental results.In view of the above problems,this paper proposes to use hybridization chain reaction(HCR)instead of PCR to amplify DNA fragments.This method is an enzyme-free isothermal nucleic acid amplification technology,which is simple,mild and does not require expensive equipment.In addition,students can deepen their understanding and application of the principle of HCR and base pairing by guiding them to design DNA sequences.HCR-based DNA fragment amplification avoids the use of expensive instruments and reagents,reduces the experimental cost,simplifies the experimental operation,and improves the success rate of the experiment.

关 键 词：DNA扩增 聚合酶链式反应 杂交链式反应 琼脂糖凝胶电泳 实验改进 

分 类 号：G642[文化科学—高等教育学] O658.9-4[文化科学—教育学]