A cost-effective RNA extraction technique from animal cells and tissue using silica columns

© 2017 by the author. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution 4.0 International License ( https://creativecommons.org/licenses/by/4.0/ )

Download PDF

Cite

XML

HTML

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.

Keywords

cell culture

extraction

RNA

silica

tissue

References

1. Alberts B (2002) From RNA to protein. In: Alberts B, Johnson A, Lewis J, Raff M, Roberts K, Walter P. Molecular Biology of the Cell. New York: Garland Science. Pp. 229-374.
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.

Previous article in this issue

Next article in this issue

Journal of Biological Methods, Electronic ISSN: 2326-9901 Print ISSN: TBA, Published by POL Scientific

Publisher's Core Philosophy

We are committed to support the scientific community by publishing impactful research and enhancing communication among scientists. At POL Scientific, we are continuously looking for ways to accelerate scientific progress and to strive for transparency and open communication, making knowledge freely accessible without barrier.

171 Skyview St. San Francisco, CA 94131 U.S.A.

info@polscientific.com