Abstract
Isolation of prokaryotic mRNA by the poly(dT) method has been difficult, primarily due to the great instability of the poly(A) sequence in its mRNA. We developed a simple method to remove rRNA from total RNA ofStaphylococcus aureus by cloning a PCR-amplifiedS. aureus rRNA gene fragment into a plasmid, and then synthesizing biotin-labeled antisense rRNA to subtract rRNA. By using this method,S. aureus rRNA is significantly reduced and mRNA is enriched. This method may be used to prepare prokaryotic mRNA for many molecular biology applications.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Cao, G. and Sarkar, N. (1992) Poly(A) RNA inEscherichia coli: nucleotide sequence at the junction of the lpp transcript and the polyadenylate moiety.Proc. Natl. Acad. Sci. USA 89, 7546–7550.
Robinson, K. A., Robb, F. T., and Schreier, H. J. (1994) Isolation of maltose-regulated genes from the Hyperthermophilic archaeum,Pyrococcus furiosus, by subtractive hybridization.Gene 148, 137–141.
Plum, G. and Clark-Curtiss J. E. (1994) Induction ofMycobacterium avium gene expression following phagocytosis by human macrophages.Infect. Immun. 62, 476–483.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Su, C., Sordillo, L.M. A simple method to enrich mRNA from total prokaryotic RNA. Mol Biotechnol 10, 83–85 (1998). https://doi.org/10.1007/BF02745865
Issue Date:
DOI: https://doi.org/10.1007/BF02745865