Synthesis and purification of horseradish peroxidase-labeled oligonucleotides for tyramide-based fluorescence in situ hybridization |
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| Authors: | R.P.M. van Gijlswijk M.P.C. van de Corput V. Bezrookove J. Wiegant H. J. Tanke A. K. Raap |
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| Affiliation: | (1) Laboratory for Cytochemistry and Cytometry, Department of Molecular Cell Biology, Leiden University Medical Center, Wassenaarseweg 72, 2333 AL Leiden, The Netherlands Tel.: +31-71-5276187, Fax: +31-71-5276180, NL |
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| Abstract: | A method is presented to conjugate horseradish peroxidase (HRP) to oligodeoxynucleotides for fluorescence in situ hybridization assays employing tyramide signal amplification (TSA). HRP is covalently bound to the oligonucleotide by thiol ether linkage and purified by high-performance liquid chromatography. With TSA detection, a single HRP-labeled oligonucleotide probe is sufficient for in situ detection of clustered DNA repeat sequences with a degree of repetition between 20 and 50. Accepted: 6 December 1999 |
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