Protocol optimization for simultaneous DNA and RNA co-extraction from single hard tick specimens
Hard 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...
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| Format: | Article |
| Language: | English |
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Elsevier
2021-01-01
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| Series: | MethodsX |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2215016121001084 |
| _version_ | 1819099482044760064 |
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| author | Alessandra Cafiso Giulia Chiappa Camilla Luzzago Anita Koni Daniele Bonato Xhelil Koleci Chiara Bazzocchi |
| author_facet | Alessandra Cafiso Giulia Chiappa Camilla Luzzago Anita Koni Daniele Bonato Xhelil Koleci Chiara Bazzocchi |
| author_sort | Alessandra Cafiso |
| collection | DOAJ |
| description | Hard 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. |
| first_indexed | 2024-12-22T00:47:34Z |
| format | Article |
| id | doaj.art-059f17a84fb9403b97dea990b17b291f |
| institution | Directory Open Access Journal |
| issn | 2215-0161 |
| language | English |
| last_indexed | 2024-12-22T00:47:34Z |
| publishDate | 2021-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | MethodsX |
| spelling | doaj.art-059f17a84fb9403b97dea990b17b291f2022-12-21T18:44:31ZengElsevierMethodsX2215-01612021-01-018101315Protocol optimization for simultaneous DNA and RNA co-extraction from single hard tick specimensAlessandra Cafiso0Giulia Chiappa1Camilla Luzzago2Anita Koni3Daniele Bonato4Xhelil Koleci5Chiara Bazzocchi6Department of Veterinary Medicine, Università degli Studi di Milano, Lodi, Italy; Corresponding author.Department of Veterinary Medicine, Università degli Studi di Milano, Lodi, ItalyDepartment of Veterinary Medicine, Università degli Studi di Milano, Lodi, Italy; Coordinated Research Center ''EpiSoMI'', Università degli Studi di Milano, Milano, ItalyDepartment of Veterinary Public Health, Faculty of Veterinary Medicine, Agricultural University of Tirana, Tirana, AlbaniaDepartment of Veterinary Medicine, Università degli Studi di Milano, Lodi, ItalyDepartment of Veterinary Public Health, Faculty of Veterinary Medicine, Agricultural University of Tirana, Tirana, AlbaniaDepartment of Veterinary Medicine, Università degli Studi di Milano, Lodi, Italy; Coordinated Research Center ''EpiSoMI'', Università degli Studi di Milano, Milano, Italy; Pediatric Clinical Research Center ''Romeo ed Enrica Invernizzi”, Università degli Studi di Milano, Milano, ItalyHard 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.http://www.sciencedirect.com/science/article/pii/S2215016121001084DNA/RNA co-extraction from hard tick specimens |
| spellingShingle | Alessandra Cafiso Giulia Chiappa Camilla Luzzago Anita Koni Daniele Bonato Xhelil Koleci Chiara Bazzocchi Protocol optimization for simultaneous DNA and RNA co-extraction from single hard tick specimens MethodsX DNA/RNA co-extraction from hard tick specimens |
| title | Protocol optimization for simultaneous DNA and RNA co-extraction from single hard tick specimens |
| title_full | Protocol optimization for simultaneous DNA and RNA co-extraction from single hard tick specimens |
| title_fullStr | Protocol optimization for simultaneous DNA and RNA co-extraction from single hard tick specimens |
| title_full_unstemmed | Protocol optimization for simultaneous DNA and RNA co-extraction from single hard tick specimens |
| title_short | Protocol optimization for simultaneous DNA and RNA co-extraction from single hard tick specimens |
| title_sort | protocol optimization for simultaneous dna and rna co extraction from single hard tick specimens |
| topic | DNA/RNA co-extraction from hard tick specimens |
| url | http://www.sciencedirect.com/science/article/pii/S2215016121001084 |
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