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标题：Protocol optimization for simultaneous DNA and RNA co-extraction from single hard tick specimens
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作者：Alessandra Cafiso ; Giulia Chiappa ; Camilla Luzzago 等
期刊名称：MethodsX
印刷版ISSN：2215-0161
电子版ISSN：2215-0161
出版年度：2021
卷号：8
页码：1-6
DOI：10.1016/j.mex.2021.101315
语种：English
出版社：Elsevier
摘要：AbstractHard ticks are important vectors of DNA- and RNA-based infectious microorganisms, but they also host complex microbial communities in which pathogens and symbionts can interact among each other and with the arthropod host itself. Molecular investigations on ticks and their hosted microorganisms are important for human and animal health. These analyses often imply the use of both DNA and RNA, with prompt preservation of nucleic acids after collection, and safe handling in case of low-level containment. Several commercial kits are available for the co-extraction of DNA and RNA; however, cost can be a limiting factor for the choice of this method, which also require additional reagents for nucleic acids preservation before extraction. Additionally, extraction buffers provided in commercial kits do not guarantee the safe handling in case of hazardous biological material. With the aim of epidemiological screenings for tick-borne pathogens and gene expression analyses focused on the relationship among ticks and their microbial communities, an optimized protocol for DNA and RNA co-extraction from single adult hard tick specimens (Ixodidae) has been developed using TRIzolⓇLS Reagent.A method for•DNA/RNA co-extraction from adult hard tick specimens;•Safe sample handling;•Obtaining DNA and RNA simultaneously for diagnostic procedures and RNA for gene expression/transcriptomic analyses.Graphical abstractDisplay Omitted
关键词：Nucleic acids;Purification;DNA isolation;RNA isolation;Co-extraction;Tick;PCR;Reverse transcription;Coinfection, Vector-borne pathogen