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  3. Synthesis and purification of horseradish peroxidase-labeled oligonucleotides for tyramide-based fluorescence in situ hybridization

Synthesis and purification of horseradish peroxidase-labeled oligonucleotides for tyramide-based fluorescence in situ hybridization

  • Histochem Cell Biol. 2000 Mar;113(3):175-80. doi: 10.1007/s004180050436.
R P van Gijlswijk 1,  M P van de Corput,  V Bezrookove,  J Wiegant,  H J Tanke,  A K Raap
Affiliations

Affiliation

  • 1 Department of Molecular Cell Biology, Leiden University Medical Center, The Netherlands.
PMID: 10817671 DOI: 10.1007/s004180050436
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.

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