文章基本信息

标题：Laboratory Exercise to Measure Plasmid Copy Number by qPCR
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作者：Benjamin David ; Jinbei Li ; Faisal Masood 等
期刊名称：Journal of Microbiology and Biology Education
印刷版ISSN：1935-7877
电子版ISSN：1935-7885
出版年度：2021
卷号：22
期号：2
DOI：10.1128/jmbe.00125-21
语种：English
出版社：American Society for Microbiology
摘要：ABSTRACT Quantitative PCR (qPCR) has numerous applications in biology. In an educational setting, qPCR provides students an opportunity to better understand the PCR mechanism by providing both quantitative information about the reactions and also data to troubleshoot PCRs (e.g., melt curves). Here, we present a relatively short (2-h) laboratory activity to demonstrate qPCR to quantify plasmid copy number (CN) by measuring the cycle threshold ( C T ) values for a genomic gene and a plasmid gene using transformed cells as a template. The activity can be combined with additional laboratory exercises, including bacterial transformation, to create the template to be used in the qPCRs. This lab activity is ideal for undergraduate laboratory courses that include recombinant DNA technology. (This work was presented at the 2020 Biomedical Engineering Society annual meeting).
关键词：qPCR;laboratory instruction