qSanger:Quantification of Genetic Variants in Bacterial Cultures by Sanger Sequencing
作者机构：School of Life SciencesUniversity of WarwickCoventryUK. De Novo Synthetic Biology LabI2SysBioCSIC-University of ValenciaPaternaSpain. Department of BiotechnologyChemistry and PharmacyUniversity of SienaSienaItaly. Centre for Applied Entomology and ParasitologySchool of Life SciencesKeele UniversityKeeleUK.
出 版 物：《BioDesign Research》 (生物设计研究（英文）)
年 卷 期：2023年第5卷第1期
页 面：49-57页
学科分类：0710[理学-生物学] 0831[工学-生物医学工程（可授工学、理学、医学学位）] 1203[管理学-农林经济管理] 081704[工学-应用化学] 07[理学] 08[工学] 0817[工学-化学工程与技术] 0903[农学-农业资源与环境] 070303[理学-有机化学] 0703[理学-化学] 0836[工学-生物工程] 0713[理学-生态学]
基 金：the Ministerio de Ciencia e Innovacion GeneCircuits++PID2020-118436GB-I00(A.J.) BBSRC BB/P020615/1(A.J.) EU grants EVOPROG 610730 and PhotoSynH2 HORIZON-EIC-2021-PATHFINDERCHALLENGES(grant agreement 101070948)(A.J.) School of Life Sciences departmental allocation,Keele University(R.G.)
主 题：neutral consuming transformed
摘 要：Genetic variations such as mutations and recombinations arise spontaneously in all cultured *** it is possible to identify nonneutral mutations by selection or counterselection,the identification of neutral mutations in a heterogeneous population usually requires expensive and time-consuming methods such as quantitative or droplet polymerase chain reaction and high-throughput *** mutations could even become dominant under changing environmental conditions enforcing transitory selection or *** propose a novel method,which we called qSanger,to quantify DNA using amplitude ratios of aligned electropherogram peaks from mixed Sanger sequencing *** expressing enhanced green fluorescent protein and mCherry fluorescent markers were used to validate qSanger both in vitro and in cotransformed Escherichia coli via quantitative polymerase chain reaction and fluorescence *** show that qSanger allows the quantification of genetic variants,including single-base natural polymorphisms or de novo mutations,from mixed Sanger sequencing reads,with substantial reduction of labor and costs compared to canonical approaches.
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