Abstract
Primed in situ (PRINS) labeling has become an alternative to in situ hybridization (ISH) for the localization of nucleic acid sequences in cell (1–4) and tissue preparations (5; see also. Chapter 5)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 (3,6,7). The detection limit of the PRINS technique appears to be on the order of low-copy sequences (3,8.
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References
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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). Bright-Field Microscopic Detection of Oligonucleotide PRINS-Labeled DNA in Chromosome Preparations. In: Gosden, J.R. (eds) PRINS and In Situ PCR Protocols. Methods in Molecular Biology™, vol 71. Humana Press. https://doi.org/10.1385/0-89603-395-3:13
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DOI: https://doi.org/10.1385/0-89603-395-3:13
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