Abstract
Since the advent of polymerase chain reaction (PCR), it has proven possible to clone complete genes, or parts of these, without having to construct the usually obligatory cDNA library. PCR cloning, however, is not without its own problems and limitations, such as the fidelity of the cloned sequence (1) and the fact that prior knowledge of the target gene sequence is required. It is for these reasons that the traditional cDNA library still has a valuable role to play in the isolation of any new gene.
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References
Tindall, K. R., and Kunkel, T. A. (1988) Fidelity of DNA synthesis by the Thermus aquaticus DNA polymerase. Biochemistry 27, 6008–6013.
Williams, J. G. (1981) The preparation and screening of a cDNA clone bank, in Genetic Engineering, vols. 1 and 2 (Williamson, R., ed.), Academic, NY, pp. 2–55.
St. John, T. and Davis, R. W. (1979) Isolation of galactose-inducible DNA sequences from Saccharomyces cerevisiae by differential plaque filter hybridisation. Cell 16, 443–452.
Zimmerman, C. R., Orr, W. C., Lecleic, R. F., Barnard, E. C., and Timberlake, W. E. (1980) Molecular cloning and selection of genes regulated in Aspergillus development. Cell 21, 709–715.
Hedrick, S. M., Cohen, D. I., Nielsen, E. A., and Davis, M. M. (1984) Isolation of cDNA clones encoding T-cell specific membrane associated proteins. Nature 308, 149–153.
Boll, W., Fujiswa, J. I., Niemi, I., and Wiessmann, C. (1986) A new approach to high sensitivity differential hybridisation. Gene 50, 41–53.
Duguid, J. R., Rohwer, R. G., and Seed, B. (1988) Isolation of cDNAs of scrapie modulated RNAs by subtractive hybridisation. Proc. Natl. Acad. Sci. USA 85, 5738–5742.
Travis, G. H. and Sutcliffe, J. G. (1988) Phenol emulsion-enhanced DNA driven subtractive cDNA cloning: isolation of low abundance monkey cortex specific mRNAs. Proc Natl. Acad. Sci. USA 85, 1696–1700.
Sive, H. I. and St. John, T. (1988) A simple subtractive hybridisation technique employing photoactivatable biotin and phenol extraction. Nucleic Acids Res. 16, 10,937.
Rubenstein, J. L. R., Brice, A. E. J., Ciaranello, R. D., Denney, D., Porteus, M. H., and Usdin, T. B. (1990) Subtractive hybridisation system using single stranded phagemids with directional inserts. Nucleic Acids Res. 18, 4833–4842.
Timblin, C., Battey, J., and Kuehl, W. M. (1990) Application of PCR technology to subtractive cDNA cloning: identification of genes expressed specifically in murine plasmacytoma cells. Nucleic Acids Res. 18, 1587–1593.
Hara, E., Kato, T., Nakada, S., Sekiya, S., and Oda, K. (1991) Subtractive cloning using oligo (dT)30 latex and PCR. Isolation of cDNA clones specific to undifferentiated embryonic carcinoma cells. Nucleic Acids Res. 19, 7097–7104.
Liang, P. and Pardee, A. B. (1992) Differential display of eukaryotic mRNA by means of the polymerase chain reaction. Science 257, 967–971.
Schweinfest, C. W. and Papas, T. (1992) Subtraction hybridisation: an approach to the isolation of genes differentially expressed in cancer and other biological systems: A review. Int. J. Oncol. 1, 499–506.
Hampson, I. N., Hampson, L., Cowling, G. J., and Dexter, T. M. (1992) Chemical cross-linking subtraction (CCLS): a new method for the generation of subtractive hybridisation probes. Nucleic Acids Res. 20, 2899.
Hartley, J. A., Berardini, M., Ponti, M., Gibson, N. W., Thompson, A. S., Thusrston, D. E., Hoey, B. M., and Butler, J. (1991) DNA cross-linking and sequence selectivity of Aziridinylbenzoquinones: a unique reaction of 5′GC3′ sequences with 2,5-diaziridmyl-1-4-benzoquinone upon reduction. Biochemistry 30, 11,714–11,724.
Feinberg, A. P. and Vogelstein, B. (1983) A technique for radiolabelling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 132, 6–13.
Favoloro, J. R., Treisman, R., and Kamen, R. (1980) Transcription maps of polyoma virus specific RNA: analysis of two dimensional nuclease S1 gel mapping. Methods Enzymol. 65, 718.
Glisin, V., Crkvenjakov, R., and Byus, C. (1974) RNA isolated by caesium chloride centrifugation. Biochemistry 13, 2633.
Chomczynski, P. and Sacchi, N. (1987) Single step method of RNA isolation by guanidinium sothiocyanatephenol-chloroform extraction. Anal. Biochem. 162, 156–159.
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© 1997 Humana Press Inc., Totowa, NJ
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Hampson, I.N., Hampson, L. (1997). Chemical Crosslinking Subtraction (CCLS). In: Boultwood, J. (eds) Gene Isolation and Mapping Protocols. Methods in Molecular Biology™, vol 68. Humana Press. https://doi.org/10.1385/0-89603-482-8:221
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DOI: https://doi.org/10.1385/0-89603-482-8:221
Publisher Name: Humana Press
Print ISBN: 978-0-89603-482-2
Online ISBN: 978-1-59259-554-9
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