12/3/2023 0 Comments Northern blot rna![]() So don’t let the idea of working with RNA scare you. There are numerous RNA isolation kits on the market now, and lots of RNAse inhibitors available. I think that Northern blots got a bad rap from the days when working with RNA was as hard as it got in molecular biology. And the naming tradition just continued with Western blots. So when invented the RNA method was named “Northern blot” as an homage to the original nucleic acid detection method. ![]() Northern Blots are done the same way as Southern blots but RNA is detected instead of DNA. A typical Southern blot experiment goes as follows: 1) run a DNA gel, 2) transfer gel contents onto a membrane, 3) hybridize said membrane with radioactive DNA probe, 4) wash off unbound radioactive probe, and 5) detect radioactive probe. Southern blots are a method of detecting DNA and named after the surname of their inventor Ed Southern. Northern Blots are named after their big brother: The Southern blot. However, because of its unpopular nature finding someone that can teach you about how to do a good Northern Blot can be hard. So sometimes Northern Blots are a necessary evil. Yet, qRT-PCR is prone to false positives and negatives, and reviewers may require Northern Blot confirmation of your qRT-PCR results. Northern Blots fell out of favor for two reasons: The perceived difficulty of working with RNA and because most people don’t like working with radioactivity. After all, it is more common these days to detect and quantify RNA with quantitative real time (qRT)-PCR than with Northern Blots. Biochemistry 6, 3650–3653.You might think Northern Blots are an old-fashioned technique. (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. (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. (1987) Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. 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. (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. (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.
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