RNA Isolation of Dengue Virus Type 2 with Different Precipitation Solvents : Methanol, Chloroform, and 2-Isopropanol.
Downloads
Dengue virus distributed in tropical and subtropical regions in the world. DENV viruses are transmitted between humans primarily by Aedes aegypti and Aedes albopictus mosquitoes and are endemic in most areas in which the vectors occur. Four serotypes of dengue virus are DENV-1, DENV-2, DENV-3 and DENV-4. DENV-2 is comprised of six genotypes. Ethanol precipitation is a commonly used technique for concentrating and de-salting nucleic acids (DNA or RNA) preparations in aqueous solution. RNA isolation by combining Guanidinium thiocyanate and phenol reported has been reported. In this report, we investigated RNA isolation from DENV-2 using QIAamp Mini Kit with 2-Isopropanol, Methanol, Chloroform precipitation solvent. Electrophoregram showed DNA band as the result of RNA isolation with methanol and 2-isopropanol are produced quite well. Dna band of the of RNA isolation with chloroform solvent has the lowest intensity than methanol and 2-isopropanol. This study showed that methanol and 2-isopropanol can used as precipitation solvent for isolating RNA.
Simmons, C.P., Farrar, J.J., Nguyen v, V., Wills, B., 2012. Dengue. N. Engl. J. Med. 366, 1423-1432.
Johnson , B.W., Russell, B. J., Lanciotti, Robert S., 2005. Serotype-Specific Detection of Dengue Viruses in a Fourplex Real-Time Reverse Transcriptase PCR Assay, vol.43 no.10 4977-4983.
WHO, 2017. Epidemiology Unit, Ministry of Health: Dengue update, 16 November 2017. https://reliefweb.int/report/sri-lanka/epidemiology-unit-ministry-health-dengue-update-16-november-2017.
Halstead, S.B., 2007. Dengue. Lancet 370, 1644–1652.
Halstead SB. Dengue Virus-mosquito Interactions. Annu Rev Entomol. 2008 Jan;53(1):273–91.
Weaver, S.C., and Vasilakis, Nikos., 2009, Molecular Evolution of Dengue Viruses: Contributions of Phylogenetics to Understanding the History and Epidemiology of the Preeminent Arboviral Disease, 523–540.
Henson, J.M. , French, R.C., 1993, The Polymerase Chain Reaction and Plant Disease Diagnosis, DOI: 10.1146/annurev.py.31.090193.000501.
Chomczynsk, Piotr., Sacchi, Nicoletta., 1986. Single-Step Method of RNA Isolation by Acid Guanidinium Thiocyanate-Phenol-Chloroform Extraction, 156- 159.
Lanciotti,R.S., Calisher,C.H.,Gubler,D.J.,Chang, G.J., Vorndam,A.V., 1992. Rapid Detection and Typing of Dengue Viruses from Clinical Samples by Using Reverse Transcriptase-Polymerase Chain Reaction Vol. 30, No. 3, 545-551.
Hery, 2014. Analisis DNA.
http://hery-irawan- fpk11.web.unair.ac.id/artikel_detail-107839-Umum-Analis%20DNA.html.
Surzycki, Stefan., 2000, Isolation and Purification of RNA, pp 119-144.
Oswald,Nick., 2007. Basics how ethanol precipitation of dna and rna works. https://bitesizebio.com/253/the-basics-how-ethanol-precipitation-of-dna-and-rna-works.
Ahamed S.F., Vivek, Rosario., Kotabagi, Shalini., Nayak,Kaustuv., Chandele, Anmol, Kaja, M.K., Shet, Anita., 2017. Enhancing the sensitivity of Dengue virus serotype detection by RT-PCR among infected children in India.
Kotaki, T., Yamanaka, A.,Mulyatno, K.C., Churrotin, S., Labiqah, A., Sucipto, T.H., Soegijanto, S., Kameoka, M., Konishi, E., 2014. Continuous dengue type 1 virus genotype shifts followed by co-circulation, clade shifts and subsequent disappearance in Surabaya, Indonesia, 2008–2013. Infect. Genet. Evol. 28, 48–54.
Kotaki, T., Yamanaka, A.,Mulyatno, K.C., Churrotin, S., Labiqah, A., Sucipto, T.H., Labiqah, Amaliah., Ahwanah, N.L.F Soegijanto, S., Kameoka, M., Konishi, E., 2016. Divergence of the dengue virus type 2 Cosmopolitan genotype associated with two predominant serotype shifts between 1 and 2 in Surabaya, Indonesia, 2008–2014. Infection, Genetics and Evolution 37 , 88–93.
Manning, Kenneth., 1990. Isolation of Nucleic Acids from Plants by Differential Solvent Precipitation. Analytical Biochemistry 195, 45-50 (19%).
Ma, Lixin., Jones, C.T., Groesch, T.D., Kuhn, R. J., Post , C.B., 2003. Solution structure of dengue virus capsid protein reveals another fold.
Su,Xing.,Gibor, Aharon., 1988. A Method for RNA Isolation from Marine Macro-Algae, Analytical Biochemistry 174,650-657(1988).
Amanda, U.D. and Cartealy, I.C. 2015. Total RNA isolation from the mesocarp of oil palm ( Elaeis guineenssis Jacq. Var Tenera) fruits. PROS SEM NAS MASY BIODIV INDON, vol 1, pages : 171-176.
Farrell, RE. 2005. RNA methodologies : A laboratory guide for isolation and characterization. 3rd ed. Elsevier Academic Press, Burlington.
COPYRIGHT NOTICE
1. By submitting the article to Jurnal Kimia Riset (JKR), the author has agreed to transfer some of the copyrights to the publisher of the research chemistry journal, Universitas Airlangga, indicated in the Copyright Transfer Agreement.
2. Authors still retain significant rights to use and share their own published articles for non-commercial purposes subject to Creative Commons Attribution-NonComercial 4.0 International License
3. All publications (printed/electronic) are open access for educational purposes, research, library, and other non-commercial purposes. Besides the purposes mentioned above, the editorial board is not responsible for copyright violations.
