Preservation of DNA and RNA for Molecular Biological Diagnosis
Preservation of DNA and RNA for Molecular Biological Diagnosis
Keywords:
Genetic material, DNA, RNA, Genetic material degradation, Genetic material preservationAbstract
DNA and RNA are genetic information codes that play a crucial role in the regulation and transmission of hereditary traits in living organisms. These genetic materials can be degraded by various environmental factors such as temperature, humidity, UV radiation, and contamination with DNase and RNase enzymes in laboratory materials and equipment. Degradation of DNA and RNA can interfere with
molecular diagnostic procedures. Preventing the degradation of genetic materials can be accomplished by chemical methods, such as the use of antioxidants, chelating agents, pH, physical processes such as storing samples at -80°C and avoiding exposure to UV light, and biological methods such as using sterile materials and equipment and preventing microbial activity. These processes are suitable for applications in preserving DNA, RNA, and biological samples for molecular analysis in medical laboratories.
References
Kawane K, Motani K, Nagata S. DNA degradation and its defects. Cold Spring Harb Perspect Biol 2014; 6(6): a016394. (14 pages).
Bhoyar L, Mehar P, Chavali K. An overview of DNA degradation and its implications in forensic caseworks. Egypt J Forensic Sci 2024; 14(1): 15. (19 pages).
Camacho-Sanchez M, Burraco P, Gomez-Mestre I, Leonard JA. Preservation of RNA and DNA from mammal samples under field conditions. Mol Ecol Resour 2013; 13(4): 663–73.
Kasu M, Ristow PG, Burrows AM, Kuplik Z, Gibbons MJ, D'Amato ME. Novel buffer for long-term preservation of DNA in biological material at room temperature. Biotechniques 2024; 76(8): 357–70.
żarczyńska M, żarczyński P, Tomsia M. Nucleic acids persistence—benefits and limitations in forensic genetics. Genes 2023; 14(8): 1643. (26 pages).
Keller W, Crouch R. Degradation of DNA RNA hybrids by ribonuclease H and DNA polymerases of cellular and viral origin. Proc Natl Acad Sci USA 1972; 69(11): 3360–4.
An R, Dong P, Komiyama M, Pan X, Liang X. Inhibition of nonenzymatic depurination of nucleic acids by polycations. FEBS Open Bio 2017; 7(11): 1707–14.
Wikipedia. Depurination. [online]. 2021; [cited 2024 Dec 22]; [2 screens]. Available from: URL: https://en.wikipedia.org/wiki/Depurination.
Wikipedia AP site. [online]. 2020; [cited 2024 Dec 22]; [3 screens]. Available from: URL: https://en.wikipedia.org/wiki/AP_site.
Houseley J, Tollervey D. The many pathways of RNA degradation. Cell 2009; 136(4): 763–76.
Wikipedia. Antioxidant. [online]. 2019; [cited 2024 Dec 22]; [11 screens]. Available from: URL: https://en.wikipedia.org/wiki/Antioxidant.
Nkuna R, Ijoma GN, Matambo TS. Applying EDTA in chelating excess metal ions to improve downstream DNA recovery from mine tailings for long-read amplicon sequencing of acidophilic fungi communities. J Fungi 2022; 8(5): 419. (18 pages).
Wikipedia. Category: chelating agents. [online]. 2020; [cited 2024 Dec 23]. Available from: URL: https://en.wikipedia.org/wiki/Category:Chelating_agents.
Wikipedia. Sodium dodecyl sulfate. [online]. 2019; [cited 2024 Dec 23]; [4 screens]. Available from: URL: https://en.wikipedia.org/wiki/Sodium_dodecyl_sulfate.
Wikipedia. Dithiothreitol. [online]. 2019; [cited 2024 Dec 23]; [4 screens]. Available from: URL: https://en.wikipedia.org/wiki/Dithiothreitol.
Wikipedia. Pyrimidine dimer. [online]. 2019; [cited 2024 Dec 27]; [5 screens]. Available from: URL: https://en.wikipedia.org/wiki/Pyrimidine_dimer.
Minas K, McEwan NR, Newbold CJ, Scott KP. Optimization of a high-throughput CTAB-based protocol for the extraction of qPCR-grade DNA from rumen fluid, plant and bacterial pure cultures. FEMS Microbiol Lett 2011; 325(2): 162–9.
Beknazarova M, Millsteed S, Robertson G, Whiley H, Ross K. Validation of DESS as a DNA preservation method for the detection of Strongyloides spp. in canine feces. Int J Environ Res Public Health 2017; 14(6): 624. (7 pages).
Jones A, Fujiyama C, Smith K. RNA purification and quantification methods. Methods Mol Med 2001; 57: 241–53.
Claeys M, Al Obaidi S, Bruyland K, Vandecandelaere I, Vandesompele J. Assessment of DNA/RNA defend pro: an inactivating sample collection buffer for enhanced stability, extraction-free PCR, and rapid antigen testing of nasopharyngeal swab samples. Int J Mol Sci 2024; 25(16): 9097. (9 pages).
Shedlovskiy D, Shcherbik N, Pestov DG. One-step hot formamide extraction of RNA from Saccharomyces cerevisiae. RNA Biol 2017; 14(12): 1722–6.
Huang YH, Leblanc P, Apostolou V, Stewart B, Moreland RB. Comparison of Milli-Q PF Plus water to DEPC-treated water in the preparation and analysis of RNA. Biotechniques 1995; 19(4): 656–61.
Mordant A, Kleiner M. Evaluation of sample preservation and storage methods for metaproteomics analysis of intestinal microbiomes. Microbiol Spectr 2021; 9(3): e0187721. (14 pages).
Dankai W, Khunamornpong S, Siriaunkgul S, Soongkhaw A, Aithin P, Lekawanvijit S. An evaluation of phosphate buffer saline as an alternative liquid-based medium for HPV DNA detection. Asian Pac J Cancer Prev 2021; 22(11): 3441–5.
Jensen M, Wippler J, Kleiner M. Evaluation of RNAlater as a field-compatible preservation method for metaproteomic analyses of bacterium-animal symbioses. Microbiol Spectr 2021; 9(2): e0142921. (14 pages).
Thermo Fisher Scientific. RNAlater solutions for RNA stabilization and storage. [online]. 2024; [cited 2024 Dec 29]; [5 screens]. Available from: URL: https://www.thermofisher.com/th/en/home/brands/product-brand/rnalater.html.
Panda BB, Meher AS, Hazra RK. Comparison between different methods of DNA isolation from dried blood spots for determination of malaria to determine specificity and cost effectiveness. J Parasit Dis 2019; 43(3): 337–42.
Sorensen A, Berry C, Bruce D, Gahan ME, Hughes-Stamm S, McNevin D. Direct-to-PCR tissue preservation for DNA profiling. Int J Legal Med 2016; 130(3): 607–13.
MySkinRecipes. TRIS HCl (Tris Hydrochloride). [online]. 2024; [cited 2024 Dec 29]. Available from: URL: https://www.myskinrecipes.com/shop/th/biological-buffers/35430-tris-hcl-tris-hydrochloride.html.
Wikipedia. TAE buffer. [online]. 2019; [cited 2024 Dec 29]; [2 screens]. Available from: URL: https://en.wikipedia.org/wiki/TAE_buffer.
Wikipedia. TBE buffer. [online]. 2020; [cited2024 Dec 29]; [2 screens]. Available from: URL: https://en.wikipedia.org/wiki/TBE_buffer.
Wikipedia. SSC buffer. Securly - Geolocation sharing. [online]. 2025; [cited 2025 Mar 29]; [1 screen]. Available from: URL: https://en.wikipedia.org/wiki/SSC_buffer.
Smith S, Morin PA. Optimal storage conditionsfor highly dilute DNA samples: a role for trehalose as a preserving agent. J Forensic Sci 2005; 50(5): 1101–8.
Habibi P, Ostad SN, Heydari A, Aliebrahimi S, Montazeri V, Foroushani AR, et al. Effect of heat stress on DNA damage: a systematic literature review. Int J Biometeorol 2022; 66(11): 2147–58.
Wikipedia. Tris. [online]. 2020; [cited 2024 Dec 27]; [4 screens]. Available from: URL: https://en.wikipedia.org/wiki/Tris.
Goetz AE, Wilkinson M. Stress and the nonsense-mediated RNA decay pathway. Cell Mol Life Sci 2017; 74(19): 3509-31.
Fowler EV, Starkie ML, Blacket MJ, Mayer DG, Schutze MK. Effect of temperature and humidity on insect DNA integrity evaluated by real-time PCR. J Econ Entomol 2024; 117(5): 1995–2002.
National Center for Biotechnology Information (NCBI). Pyrimidine Dimers [MeSH Terms]. MeSH Database. [online]. 2024; [cited 2024 Dec 27]. Available from: https://www.ncbi.nlm.nih.gov/mesh?Cmd=DetailsSearch&Term=%22Pyrimidine+Dimers%22%5BMeSH+Terms%5D.
Henry E, Charalambous E, Betsou F, Mathieson W. Implementing routine monitoring for nuclease contamination of equipment and consumables into the quality Management system of a laboratory. Heliyon 2024; 10(2): e24603. (11 pages).
Qamar W, Khan MR, Arafah A. Optimization of conditions to extract high quality DNA for PCR analysis from whole blood using SDS-proteinase K method. Saudi J Biol Sci 2017; 24(7): 1465–9.
Sohrabipour S, Muniz VS, Sharma N, Dwivedi DJ, Liaw PC. Mechanistic studies of DNase I activity: impact of heparin variants and PAD4. shock 2021; 56(6): 975–87.
Xi Z, Wang Z, Sarafianos SG, Myshakina NS, Ishima R. Determinants of active-site inhibitor interaction with HIV-1 RNase H. ACS Infec Dis 2019; 5(11): 1963–74.
Latham KE, Miller JJ. DNA recovery and analysis from skeletal material in modern forensic contexts. Forensic Sci Res 2018; 4(1): 51–9.
Kirkland PD, Frost MJ. The impact of viral transport media on PCR assay results for the detection of nucleic acid from SARS-CoV-2. Pathology 2020; 52(7): 811–4.
Nunes JS, Pimentel-Vera LN, Silva SB, de Bem Prunes B, Rados PV, Visioli F. Comparison of different DNA preservation solutions for oral cytological samples. Arch Oral Biol 2024; 158: 105867. (7 pages).
Tuttle RM, Waselenko JK, Yosseffi P, Weigand N, Martin RK. Preservation of nucleic acids for polymerase chain reaction after prolonged storage at room temperature. Diagn Mol Pathol 1998; 7(6): 302–9.
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