Abstract
Primed in situ (PRINS) labeling has become an alternative to in situ hybridization (ISH) for the localization of nucleic acid sequences in cell preparations (1–4). In the PRINS method, an unlabeled primer (restriction fragment, PCR product, or oligonucleotide) is annealed to its complementary target sequence in situ. The primer serves as an initiation site for in situ chain elongation using a thermostable DNA polymerase and labeled nucleotides, which can be detected directly by fluorescence microscopy, such as fluorochrome-labeled dNTPs, or indirectly using, e.g., biotin- or digoxigenin-dUTP and the application of fluorochrome-conjugated avidin or antibody molecules (3,5,6). The detection limit of the PRINS technique appears to be in the order of low-copy sequences (3,7).
References
Bains, M. A., Agarwal, R., Pringle, J. H., Hutchinson, R. M., and Lauder, I. (1993) Flow cytometric quantitation of sequence-specific mRNA in hemapoietic cell suspensions by primer-induced in situ (PRINS) fluorescent nucleotide labeling. Exp. Cell Res. 208, 321–326.
Koch, J., Mogensen, J., Pedersen, S., Fischer, H., Hindkjær, S., Kölvraa, S., and Bolund, L. (1992) Fast one-step procedure for the detection of nucleic acids in situ by primer-induced sequence-specific labeling with fluorescein-12-dUTP. Cytogenet. Cell Genet. 60, 1–3.
Gosden, J. and Lawson, D. (1994) Rapid chromosome identification by oligonucleotide-primed in situ DNA synthesis (PRINS). Hum. Mol. Genet. 3, 931–936.
Pellestor, F., Girardet, A., Lefort, G., Andrea, B., and Charlieu, J. P. (1995) PRINS as a method for rapid chromosomal labeling of human spermatozoa. Mol. Reprod. Dev. 40, 333–337.
Hindkjær, J., Koch, J., Terkelsen, C., Brandt, C. A., Kølvraa, S., and Bolund, L. (1994) Fast, sensitive multicolor detection of nucleic acids in situ by primed in situ labeling (PRINS). Cytogenet. Cell Genet. 66, 152–154.
Speel, E. J. M., Lawson, D., Hopman, A. H. N., and Gosden, J. (1995) Multi-PRINS: multiple sequential oligonucleotide primed in situ DNA synthesis reactions label specific chromosomes and produce bands. Hum. Genet. 95, 29–33.
Abbo, S., Dunford, R. P., Miller, T. E., Reader, S. M., and King, I. P. (1993) Primer-mediated in situ detection of the B-hordein gene cluster on barley chromosome 1H. Proc. Natl. Acad. Sci. USA 90, 11,821–11,824.
Speel, E. J. M., Lawson, D., Ramaekers, F. C. S., Gosden, J. R., and Hopman, A. H. N. (1996) Rapid brightfield detection of oligonucleotide primed in situ (PRINS) labeled DNA in chromosome preparations and frozen tissue sections. Biotechniques 20, 226–234.
Hopman, A. H. N., Van Hooren, E., Van der Kaa, C. A., Vooijs, G. P., and Ramaekers, F. C. S. (1991) Detection of numerical chromosome aberrations using in situ hybridization in paraffin sections of routinely processed bladder cancers. Mod. Pathol. 4, 503–513.
Hopman, A. H. N., Poddighe, P. J., Moesker, O., and Ramaekers, F. C. S. (1992) Interphase cytogenetics: an approach to the detection of genetic aberrations in tumours, in Diagnostic Molecular Pathology, A Practical Approach, vol. 1 (Herrington, C. S. and McGee, J. O’. D., eds.), IRL, Oxford, pp. 141–167.
Speel, E. J. M., Jansen, M. P. H. M., Ramaekers, F. C. S., and Hopman, A. H. N. (1994) A novel triple-color detection procedure for brightfield microscopy, combining in situ hybridization with immunocytochemistry. J. Histochem. Cytochem. 42, 1299–1307.
Bobrow, M. N., Harris, T. D., Shaughnessy, K. J., and Litt, G. J. (1989) Catalyzed reporter deposition, a novel method of signal amplification: Amplification to immunoassays. J. Immunol. Methods 125, 279–285.
Speel, E. J. M., Ramaekers, F. C. S., and Hopman, A. H. N. (1995) Cytochemical detection systems for in situ hybridization, and the combination with immunocytochemistry. Histochem. J. 27, 833–858.
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© 1997 Humana Press Inc, Totowa, NJ
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Speel, E.J.M., Lawson, D., Ramaekers, F.C.S., Gosden, J.R., Hopman, A.H.N. (1997). PRINS DNA Synthesis on Frozen Tissue Sections. In: Gosden, J.R. (eds) PRINS and In Situ PCR Protocols. Methods in Molecular Biology™, vol 71. Humana Press. https://doi.org/10.1385/0-89603-395-3:31
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DOI: https://doi.org/10.1385/0-89603-395-3:31
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