RNA: Methods for Preparation

Abstract

Isolation of RNA is the first critical step in analysing gene expression and viral RNA function and the cloning of genes. Isolation must free the RNA contaminants that may interfere with subsequent applications and must inactivate RNA‐degrading enzymes.

Keywords: RNA isolation; RNA extraction; ribonuclease; messenger RNA; guanidine; northern analysis

References

Aviv H and Leder P (1972) Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acid‐cellulose. Proceedings of the National Academy of Sciences of the USA 69: 1408–1412.

Chirgwin JM, Przybyla AM, MacDonald RJ and Rutter WJ (1979) Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry 18: 5294–5299.

Chomczynski P (1993) A reagent for the single‐step simultaneous isolation of RNA, DNA and proteins from cell and tissue samples. Biotechniques 15: 532–535.

Chomczynski P and Sacchi N (1987) Single‐step method of RNA isolation by acid guanidinium thiocyanate–phenol–chloroform extraction. Analytical Biochemistry 162: 156–159.

Cleaver O, Patterson KD and Kreig PA (1996) Isolation of poly‐A RNA. In: Kreig PA (ed.) A Laboratory Guide to RNA, pp. 51–64. New York: Wiley.

Further Reading

Kingston RE (1999) Preparation and analysis of RNA. In: Ausubel FM et al. (eds) Current Protocols in Molecular Biology, pp. 4.0.1–4.5.2. New York: Wiley.

Rapley R and Manning DL (eds) (1998) RNA Isolation and Characterization Protocols. Totowa, NJ: Humana Press.

Siebert PD and Larrick JW (eds) (1998) Obtaining RNA for RT‐PCR. In: Gene Cloning and Analysis by RT‐PCR, pp. 3–68. Natick, MA: Eaton Publishing.

Contact Editor close
Submit a note to the editor about this article by filling in the form below.

* Required Field

How to Cite close
Chomczynski, Piotr, and Mackey, Karol(Apr 2001) RNA: Methods for Preparation. In: eLS. John Wiley & Sons Ltd, Chichester. http://www.els.net [doi: 10.1038/npg.els.0002607]