A simple method to enrich mRNA from total prokaryotic RNA

Springer Science and Business Media LLC - Tập 10 - Trang 83-85 - 1998
Chunlei Su1, Lorraine M. Sordillo1
1Dept. of Veterinaty Science, Pennsylvannia State University, University Park

Tóm tắt

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.

Tài liệu tham khảo

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.