![]() Science 232: 1385–1389.A primary limitation of all blot hybridizations is the efficiency of hybridization between the nucleic acids on the membrane, and the labeled nucleic acids in the hybridization solution. Williams PH, Hili CB (1986) Rapid cycling populations of Brassica. Wetmur JG (1975) Acceleration of DNA renaturation rates. Wahl GM, Stern M, Stark GR (1979) Efficient transfer of large DNA fragments from agarose gels to diazobenzyloxymethyl-paper and rapid hybridization using dextran sulfate. Tautz O, Renz M (1983) An optimized freeze-squeeze method for the recovery of DNA from agarose gels. Southern EM (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. Cold Spring Harbor, NY: Cold Spring Harbor Press. Sambrook J, Fritsch EF, Maniatis T (1989) Molecular Cloning: A Laboratory Manual, 2nd ed. Rigby PWJ, Oieckman M, Rhodes C, Berg P (1977) Labelling deoxyribonucleic acid to high specific activity in vitra by nick translation with ONA polymerase I. Reed KC, Mann DA (1985) Rapid transfer of DNA from agarose gels to nylon membranes. Pichersky E, Hoffman NE, Malik VS, Bernatzky R, Tanksley SO, Szabo L, Cashmore AR (1987) The tomato Cab-4 and Cab-5 genes encode a second type of CAB polypeptides localized in Photosystem H. Milier JC, Tanksley SO (1990) Effect of different restriction enzymes, probe source, and probe length on detecting restriction fragment length polymorphism in tomato. Landry BS, Kesseli R, Leung H, Michelmore RW (1987) Comparison of restriction endonucleases and sources of probes for their efficiency in detecting restriction fragment length polymorphisms in lettuce (Lactuca sativa L.). Helentjaris T (1987) A genetic linkage map in maize based on RFLPs. Havey MJ, Muehlbauer FJ (1989) Linkages between restriction fragment length, isozyme, and morphological markers in lentil. Harnes BO, Higgins SJ, eds (1985) Nucleic Acid Hybridization: A Practical Approach. Gebhardt C, Ritter E, Oebener T, Schachtschabel U, Walkerneier B, Uhrig H, Salamini F (1989) RFLP analysis and linkage mapping in Solanum tuberosum. Plant Mol Biol Rep 4: 37–41.įeinberg AP, Vogelstein B (1984) A technique for radiolabelling ONA restriction fragments to high specific activity. Martinus Nijhoff, Oordrecht, Netherlands.īernatzky R, Tanksley SO (1986) Methods for detecting single or low copy sequences in tomato on Southern blots. In: Gelvin SB, Schilperoort RA, Verma OPS (eds) Plant Molecular Biology Manual, pp. Oxford: IRL Press.īernatzky R (1988) Restriction fragment length polymorphisms. In: Harnes BO, Higgins SJ (eds) Nucleic Acid Hybridization: A Practical Approach, pp. This process is experimental and the keywords may be updated as the learning algorithm improves.Īnderson MML, Young BO (1985) Quantitative filter hybridization. These keywords were added by machine and not by the authors. Through autoradiography, one or a few fragments can be visualized separately from the very large number of fragments that make up a complex genome. The membrane-bound DNA is then hybridized in solution to specific radio labelled DNA sequences (probes) that anneal to complementary sequences on the membrane. The fragments are transferred in a single-stranded state by treating the gel with alkali which disrupts the hydrogen bonding between the native double-stranded DNA. Briefly, the procedure involves the enzymatic cleavage of DNA with restriction endonucleases, the separation of the resultant fragments by electrophoresis through an agarose gel and the transfer of the fragments from the gel to a membrane that binds nucleic acids. ![]() At present, the technique that remains central to RFLP analysis is Southern blotting and hybridization.
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