A cost-effective RNA extraction technique from animal cells and tissue using silica columns
Main Article Content
Keywords
cell culture, extraction, RNA, silica, tissue
Abstract
Ribonucleic acid (RNA) is widely used in molecular biology assays, and some of the most common assays include: northern blotting and RT-PCR gene expression analysis. RNA is generally extracted by two methods: phenol-chloroform or commercially available silica spin column kits. Phenol-chloroform extraction is generally more economical; however, it produces hazardous byproducts, and leftover chemicals in the sample that can inhibit downstream applications. Commercial kits usually have simple set ups and short preparation time; however, they can introduce a significant expense to laboratory budgets. Here we have created a method to extract RNA using generic silica columns and readily available reagents while maintaining a high yield and purity.
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References
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2. Boom R, Sol CJ, Salimans MM, Jansen CL, Wertheim-van Dillen PM, van der Noordaa J (1990) Rapid and simple method for purification of nucleic acids. J Clin Microbiol 28: 495-503.
3. Borodina TA, Lehrach H, Soldatov AV (2003) DNA purification on homemade silica spin-columns. Analytical Biochemistry 321: 135-137. doi: 10.1016/S0003-2697(03)00403-2.
4. Gallego Romero I, Pai AA, Tung J, Gilad Y (2014) RNA-seq: impact of RNA degradation on transcript quantification. BMC Biology 12: 42. doi: 10.1186/1741-7007-12-42
5. Gonzalez-Perez I, Armas Cayarga A, Garcia de la Rosa, Iria, Josefina Gonzalez Gonzalez Y (2007) Homemade viral RNA isolation protocol using silica columns: A comparison of four protocols. Analytical Biochemistry 360: 148-150. doi: 10.1016/j.ab.2006.10.022.
6. Gordon JA (1972) Denaturation of globular proteins. Interaction of guanidinium salts with three proteins. Biochemistry 11: 1862-1870. doi: 10.1021/bi00760a021.
7. Serganov A, Nudler E (2013) A Decade of Riboswitches. Cell 152: 17-24. doi: 10.1016/j.cell.2012.12.024.
8. Sohlenius-Sternbeck A (2006) Determination of the hepatocellularity number for human, dog, rabbit, rat and mouse livers from protein concentration measurements. Toxicology in Vitro 20: 1582-1586. doi: 10.1016/j.tiv.2006.06.003.
9. Useful Numbers for Cell Culture. ThermoFisher Scientific. Cited on January 11, 2017. Available from: https://www.thermofisher.com/us/en/home/references/gibco-cell-culture-basics/cell-culture-protocols/cell-culture-useful-numbers.html.
10. Wong K, Pang H (2013) Simplifying HT RNA quality & quantity analysis. GEN 33: 17. doi: 10.1089/gen.33.2.09.
11. Xiang X, Qiu D, Hegele RD, Tan WC (2001) Comparison of different methods of total RNA extraction for viral detection in sputum. Journal of Virological Methods 94: 129-135. doi: 10.1016/S0166-0934(01)00284-1.
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Published
May 10, 2017
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How to Cite
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Escobar MD, Hunt JL. A cost-effective RNA extraction technique from animal cells and tissue using silica columns. J Biol Methods [Internet]. 2017 May 10 [cited 2024 Apr. 26];4(2):e72. Available from: https://jbmethods.org/index.php/jbm/article/view/184
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