Abstract
The purpose of this chapter is to give an overview of the different end labeling procedures. These protocols have been standardized and optimized by several biotechnology companies and are available in kits. Unless your laboratory is involved in producing large quantities of many different probes (e.g., micrograms of each probe), it is unpractical, and perhaps more expensive, to set up your own protocol. However, a good understanding of the type of probe, the location of the label (3′-, 5′-end ordistributed all along the molecule), and the number of labeled nucleotides incorporated should be considered when planning an experiment.
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References
Derbyshire, V., Freemont, P. S., Sanderson, M. R., Beese, L., Friedman, J. M., Joyce, C. M., and Steitz, T. A. (1988) Genetic and crystallographic studies of the 3′,5′-exonucleolytic site of DNA polymerase I. Science 240, 199–201.
Challberg, M.D. and Englund, P. T. (1980) Specific labeling of 3′ termini with T4 DNA polymerase. Methods Enzymol. 65, 39–43.
Deng, G. and Wu, R. (1983) Terminal transferase: use of the tailing of DNA and for in vitro mutagenesis. Methods Enzymol. 100, 96–115.
Chang, L. M. and Bollum, F. J. (1986) Molecular biology of terminal transferase. CRC Crit. Rev. Biochem. 21, 27–52.
Eschenfeldt, W.H., Puskas, R. S., and Berger, S. L. (1987) Homopolymeric tailing. Methods Enzymol. 152, 337–342.
Tu, C. and Cohen, S. (1980) 3′-End labeling of DNA with [γ-32P]cordycepin-5′-triphosphate. Gene 10, 177–183.
Vincent, C, Tchen, P., Cohen-Solal, M., and Kourilsky, P. (1982) Synthesis of 8-(2-4-dinitrophenyl 2-6 aminohexyl) amino-adenosine 5′ triphosphate: biological properties and potential uses. Nucleic Acids Res. 10, 6787–6796.
Kumar, A., Tchen, P., Roullet, F., and Cohen, J. (1988) Non radioactive labeling of synthetic oligonucleotide probes with terminal deoxynucleotidyl transferase. Anal. Biochem. 169, 376–382.
Brown, T. A., ed. (1998) Polecular Biology Labfax, I. Recombinant DNA, 2nd edit., Academic Press, San Diego.
Schmitz, G. G., Walter, T., Seibl, R., and Kessler, C. (1991) Nonradioactive labeling of oligonucleotides in vitro with the hapten digoxigenin by tailing with terminal transferase. Anal. Biochem. 192, 222–231.
Cameron, V. and Uhlenbeck, O. C. (1977) 3′-Phosphatase activity in T4 polynucleotide kinase. Biochemistry 16, 5120–5126.
Cameron, V., Soltis, D., and Uhlenbeck, O. C. (1978) Polynucleotide kinase from a T4 mutant which lacks the 3′ phosphatase activity. Nucleic Acids Res. 5, 825–833.
Sambrook, J., Fritch, E. F., and Maniatis, T. (1989) Molecular Cloning: A Laboratory Manual, 2nd edit., Cold Spring Harbor Laboratory, Cold Spring Harbor, New York.
Ausubel, F. M., Brent, R., Kingston, R. E., Moore, D. D., Seidman, J. G., Smith, J. A., and Struhl, K., eds. (1994) Current Protocols in Molecular Biology. John Wiley & Sons, New York.
Zischler, H., Nanda, I., Schafer, R., Schmid, M., and Epplen, J. T. (1989) Digoxigenated oligonucleotide probes specific for simple repeats in DNA fingerprinting and hybridization in situ. Hum. Genet. 82, 227–233.
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Hilario, E. (2002). End Labeling Procedures. In: de Muro, M.A., Rapley, R. (eds) Gene Probes. Methods in Molecular Biology, vol 179. Humana Press, Totowa, NJ. https://doi.org/10.1385/1-59259-238-4:013
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DOI: https://doi.org/10.1385/1-59259-238-4:013
Publisher Name: Humana Press, Totowa, NJ
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