Abstract
The protocol is suited for the quantitative and qualitative detection of simple repeat target DNA composed of three or fewer of the four bases A, C, G, and T. A consequence of the lacking base(s) is that such DNA can be synthesized from nucleotide mixtures containing the particular bases as dideoxynucleotides. Most genomic DNA contains all four bases and can therefore not be synthesized from such a nucleotide mixture. The combined effects of probe specificity and selective DNA synthesis from the nucleotide mixture improves the signal-to-noise ratio for such targets approximately an order of magnitude, enabling the detection of exceedingly small hybridization targets (e.g., variant telomeric repeat variants embedded in, or situated next to, the main repeat), provided that they present enough template for the DNA synthesis.
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References
Krejci, K. and Koch, J. (1998) Improved detection and comparative sizing of human chromosomal telomeres in situ. Chromosoma 107, 198–203.
Serakinci, N. and Koch, J. (1999) Detection and sizing of human telomeric repeat DNA in situ. Nat. Biotech. 17, 200–201.
Krejci, K. and Koch, J. (1999) An in situ study of variant telomeric repeats. Genomics 58, 202–206.
Serakinci, N. Krejci, K., and Koch, J. (1999) Telomeric repeat organization-a comparative in situ study between man and rodent. Cytogenet. Cell Genet. 86, 204–211.
Koch, J., Kølvraa, S., Gregersen, N., and Bolund, L. (1989) Oligonucleotide-priming methods for the chromosome-specific labeling of alpha satellite DNA in situ. Chromosoma 98, 259–265.
Kadandale, J. S., Tunca, Y., and Tharapel, A. T. (2000) Chromosomal localization of single copy genes SRY and SOX3 by primed in situ labeling (PRINS). Microb Comp Genomics 5, 71–74.
Kadandale, J. S., Wachtel, S. S., Tunca, Y., Wilroy, R. S. Jr., Martens, P. R., and Tharapel, A. T. (2000) Localization of SRY by primed in situ labeling in XX and XY sex reversal. Am. J. Med. Genet. 95, 71–74.
Therkelsen, A. J., Nielsen, A., Koch, J., Hindkjaer, J., and Kølvraa, S. (1995) Staining of Human telomeres with primed in situ labeling (PRINS). Cytogenet. Cell Genet. 68, 115–118.
Koch, J., Hindkjær, J., Kølvraa, S., and Bolund, L. (1995) Construction of a panel of chromosome specific oligonucleotide probes (PRINS-primers) useful for the identification of individual human chromosomes in situ. Cytogenet. Cell Genet. 71, 142–147.
Koch, J., Hindkjær, J., Mogensen, J., Kølvraa, S., and Bolund, L. (1991) An improved method for chromosome-specific labelling of alpha satellite DNA in situ using denatured double stranded DNA probes as primers in a PRimed IN Situ labelling (PRINS) procedure. Genet. Anal. 8, 171–178.
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Koch, J. (2006). Detection and Sizing of Telomeric and Other Simple Repeats by Dideoxy-PRINS. In: Pellestor, F. (eds) PRINS and In Situ PCR Protocols. Methods in Molecular Biology™, vol 334. Humana Press. https://doi.org/10.1385/1-59745-068-5:81
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DOI: https://doi.org/10.1385/1-59745-068-5:81
Publisher Name: Humana Press
Print ISBN: 978-1-58829-549-1
Online ISBN: 978-1-59745-068-3
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