![]() The semiporous agarose allows molecules of DNA to pass with varying degrees of ease, at a rate inversely proportional to their size. (Because the phosphate groups in DNA make the molecules negatively charged, they will migrate toward the anode in an electric field. ![]() Electrophoresis through an agarose gel then separates these fragments according to size. Purified genomic DNA is digested with a specific restriction endonuclease, which, as described above, will produce an array of differently sized DNA fragments. The presence of these sequences usually means that the gene itself is present in the genomic DNA. 17 In general, it allows one to determine whether specific nucleotide sequences in a cloned probe are present in a sample of genomic DNA. Biochemistry 6, 3650–3653.One of the most useful techniques for analyzing a gene at the level of genomic DNA is Southern blotting, named for its originator, E.M. (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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