PHOTOPROBER® Biotin: An Alternative Method for Labeling Archival DNA for Comparative Genomic Hybridization

Comparative genomic hybridization (CGH) represents a powerful method for screening the entire genome of solid tumors for chromosomal imbalances. Particularly it enabled the molecular cytogenetic analysis of archival, formalin‐fixed, paraffin‐embedded (FFPE) tissue. A well‐known dilemma, however, is...

Full description

Saved in:
Bibliographic Details
Main Authors: Dirk Korinth, Konrad Donhuijsen, Ulrike Bockmühl, Iver Petersen
Format: Article
Language:English
Published: Wiley 2004-01-01
Series:Cellular Oncology
Online Access:http://dx.doi.org/10.1155/2004/847515
Tags: Add Tag
No Tags, Be the first to tag this record!
_version_ 1832547420562522112
author Dirk Korinth
Konrad Donhuijsen
Ulrike Bockmühl
Iver Petersen
author_facet Dirk Korinth
Konrad Donhuijsen
Ulrike Bockmühl
Iver Petersen
author_sort Dirk Korinth
collection DOAJ
description Comparative genomic hybridization (CGH) represents a powerful method for screening the entire genome of solid tumors for chromosomal imbalances. Particularly it enabled the molecular cytogenetic analysis of archival, formalin‐fixed, paraffin‐embedded (FFPE) tissue. A well‐known dilemma, however, is the poor DNA quality of this material with fragment sizes below 1000 bp. Nick translation, the conventionally used enzymatic DNA labeling method in CGH, leads to even shorter fragments often below a critical limit for successful analysis. In this study we report the alternative application of non‐enzymatic, PHOTOPROBE® biotin labeling for conjugation of the hapten to the DNA prior to in situ hybridization and fluorescence detection. We analyzed 51 FFPE tumor samples mainly from the upper respiratory tract by both labeling methods. In 19 cases, both approaches were successful. The comparison of hybridized metaphases showed a distinct higher fluorescence signal of the PHOTOPROBE® samples sometimes with a discrete cytoplasm background which however did not interfere with specificity and sensitivity of the detected chromosomal imbalances. For further 32 cases characterized by an average DNA fragment size below 1000 bp, PHOTOPROBE® biotin was the only successful labeling technique thus offering a new option for CGH analysis of highly degraded DNA from archival material.
format Article
id doaj-art-325f487ae4364daa89bf52c8ceb2e8c9
institution Kabale University
issn 1570-5870
1875-8606
language English
publishDate 2004-01-01
publisher Wiley
record_format Article
series Cellular Oncology
spelling doaj-art-325f487ae4364daa89bf52c8ceb2e8c92025-02-03T06:44:43ZengWileyCellular Oncology1570-58701875-86062004-01-01265-632933410.1155/2004/847515PHOTOPROBER® Biotin: An Alternative Method for Labeling Archival DNA for Comparative Genomic HybridizationDirk Korinth0Konrad Donhuijsen1Ulrike Bockmühl2Iver Petersen3Institute of Pathology, University Hospital Charité, Berlin, GermanyInstitute of Pathology, Urban Hospital Braunschweig, GermanyClinic of Head and Neck Diseases, Hospital Fulda, GermanyInstitute of Pathology, University Hospital Charité, Berlin, GermanyComparative genomic hybridization (CGH) represents a powerful method for screening the entire genome of solid tumors for chromosomal imbalances. Particularly it enabled the molecular cytogenetic analysis of archival, formalin‐fixed, paraffin‐embedded (FFPE) tissue. A well‐known dilemma, however, is the poor DNA quality of this material with fragment sizes below 1000 bp. Nick translation, the conventionally used enzymatic DNA labeling method in CGH, leads to even shorter fragments often below a critical limit for successful analysis. In this study we report the alternative application of non‐enzymatic, PHOTOPROBE® biotin labeling for conjugation of the hapten to the DNA prior to in situ hybridization and fluorescence detection. We analyzed 51 FFPE tumor samples mainly from the upper respiratory tract by both labeling methods. In 19 cases, both approaches were successful. The comparison of hybridized metaphases showed a distinct higher fluorescence signal of the PHOTOPROBE® samples sometimes with a discrete cytoplasm background which however did not interfere with specificity and sensitivity of the detected chromosomal imbalances. For further 32 cases characterized by an average DNA fragment size below 1000 bp, PHOTOPROBE® biotin was the only successful labeling technique thus offering a new option for CGH analysis of highly degraded DNA from archival material.http://dx.doi.org/10.1155/2004/847515
spellingShingle Dirk Korinth
Konrad Donhuijsen
Ulrike Bockmühl
Iver Petersen
PHOTOPROBER® Biotin: An Alternative Method for Labeling Archival DNA for Comparative Genomic Hybridization
Cellular Oncology
title PHOTOPROBER® Biotin: An Alternative Method for Labeling Archival DNA for Comparative Genomic Hybridization
title_full PHOTOPROBER® Biotin: An Alternative Method for Labeling Archival DNA for Comparative Genomic Hybridization
title_fullStr PHOTOPROBER® Biotin: An Alternative Method for Labeling Archival DNA for Comparative Genomic Hybridization
title_full_unstemmed PHOTOPROBER® Biotin: An Alternative Method for Labeling Archival DNA for Comparative Genomic Hybridization
title_short PHOTOPROBER® Biotin: An Alternative Method for Labeling Archival DNA for Comparative Genomic Hybridization
title_sort photoprober r biotin an alternative method for labeling archival dna for comparative genomic hybridization
url http://dx.doi.org/10.1155/2004/847515
work_keys_str_mv AT dirkkorinth photoproberbiotinanalternativemethodforlabelingarchivaldnaforcomparativegenomichybridization
AT konraddonhuijsen photoproberbiotinanalternativemethodforlabelingarchivaldnaforcomparativegenomichybridization
AT ulrikebockmuhl photoproberbiotinanalternativemethodforlabelingarchivaldnaforcomparativegenomichybridization
AT iverpetersen photoproberbiotinanalternativemethodforlabelingarchivaldnaforcomparativegenomichybridization