(1967) A method for the hybridization of nucleic acid molecules at low temperature. (1977) Rates of formation and thermal stabilities of RNA:DNA and DNA:DNA duplexes at high concentrations of formamide. (1983) A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Nucleic Acids Symp Ser 10, 61–64.įeinberg, A. Application to specific modification of DNA. (1981) Photoreaction of thymidine with primary amines. Saito, I., Sugiyama, H., Furukawa, N., Matsuura, T. (1988) Rapid, reversible staining of northern blots prior to hybridization. (1980) Electrophoretic transfer of proteins and nucleic acids from slab gels to diazobenzyloxymethyl cellulose or nitrocellulose sheets. Three main blotting techniques have been de veloped and are commonly called Southern, northern, and western blotting. (1994) One-hour downward capillary blotting of RNA at neutral pH. Biochim Biophys Acta 324, 320–333.Ĭhomczynski, P., Mackey, K. (1973) Gel electrophoresis of RNA under denaturing conditions. Reijnders, L., Sloof, P., Sival, J., Borst, P. (1977) Analysis of single- and double-stranded nucleic acids on polyacrylamide and agarose gels by using glyoxal and acridine orange. (1977) RNA molecular weight determinations by gel electrophoresis under denaturing conditions, a critical reexamination. (1972) Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acid-cellulose. By using mRNA instead of total RNA, cleaner results of. This mRNA may occur in high, medium, or low abundance. Only 15 of the total RNA in the cytoplasm of a typical eukaryotic cell is mature mRNA. (1987) Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Total RNA for northern analysis will frequently result in barely detectable hybridization signals and isolation of mRNA is generally preferred. Academic, San Diego, CA.Ĭhomczynski, P., Sacchi, N. A Laboratory Guide for Isolation and Characterization. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.ĭarling, D. (1989) Molecular Cloning: A Laboratory Manual. (1994) Isolation and characterization of Ribonucleoprotein complexes, in (Higgins, S. Furthermore, biotin-labeled probes are more stable and safer for researchers and the environment than traditional probes. (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. Northern blots using RNA derived from both Arabidopsis and Oryza sativa were performed, and the results suggest that this improved method is both sensitive and efficient. (1979) Detection of specific RNAs or specific fragments of DNA by fractionation in gels and transfer to diazobenzyloxymethyl paper. (1977) Method for detection of specific RNAs in agarose gels by transfer to diazobenzyloxymethyl-paper and hybridization with DNA probes. In total, we identified 9 new sRNAs.Alwine, J. Depending on the set threshold value for fluorescence signal in our microarray approach, we identified 50–80 putative ncRNAs, 12 of which were verified by Northern blot analysis. A computational search for intergenic non-coding (nc) RNAs in the 4.6-Mb genome identified a total of 252 potential ncRNAs (including 233 putative sRNAs). the Aliivibrio salmonicida, which causes severe disease in farmed Atlantic salmon and other fishes. We have in this study significantly contributed to the discovery and verification of new sRNAs in a representative of Vibrionaceae, i.e. For example, of the hundreds of candidate sRNAs from members of the bacterial family Vibrionaceae, the function is known for only 9. ![]() However, many sRNAs remain to be identified, and the functional roles of sRNAs are known for only a small fraction. Some of the well-characterized sRNAs serve critical steps in the regulation of important cellular processes, such as quorum sensing (Qrr), iron homeostasis (RyhB), oxidative stress (OxyS), and carbon metabolism (Spot 42). Bacterial small RNAs (sRNAs) are trans-encoded regulatory RNAs that typically bind mRNAs by short-sequence complementarities and change the expression of the corresponding proteins.
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