Automated array-CGH optimized for archival formalin-fixed, paraffin-embedded tumor material.
BACKGROUND
Array Comparative Genomic Hybridization (aCGH) is a rapidly evolving technology that still lacks complete standardization. Yet, it is of great importance to obtain robust and reproducible data to enable meaningful multiple hybridization comparisons. Special difficulties arise when aCGH is performed on archival formalin-fixed, paraffin-embedded (FFPE) tissue due to its variable DNA quality. Recently, we have developed an effective DNA quality test that predicts suitability of archival samples for BAC aCGH.


METHODS
In this report, we first used DNA from a cancer cell-line (SKBR3) to optimize the aCGH protocol for automated hybridization, and subsequently optimized and validated the procedure for FFPE breast cancer samples. We aimed for highest throughput, accuracy, and reproducibility applicable to FFPE samples, which can also be important in future diagnostic use.


RESULTS
Our protocol of automated array-CGH on archival FFPE ULS-labeled DNA showed very similar results compared with published data and our previous manual hybridization method.


CONCLUSIONS
This report combines automated aCGH on unamplified archival FFPE DNA using non-enzymatic ULS labeling, and describes an optimized protocol for this combination resulting in improved quality and reproducibility.
Joosse
Simon A
SA
Division of Experimental Therapy, the Netherlands Cancer Institute, Amsterdam, The Netherlands. s.joosse@nki.nl
van Beers
Erik H
EH
Nederlof
Petra M
PM
D001331:Automation; D001707:Biopsy, Needle; D001943:Breast Neoplasms / Q000235:genetics* / Q000473:pathology; D045744:Cell Line, Tumor; D004273:DNA, Neoplasm / Q000032:analysis*; D005260:Female; D005557:Formaldehyde / Q000494:pharmacology; D006801:Humans; D009693:Nucleic Acid Hybridization / Q000379:methods*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D016612:Paraffin Embedding; D015203:Reproducibility of Results; D012494:Sampling Studies; D012680:Sensitivity and Specificity; D016707:Tissue Fixation / Q000379:methods
D004273:DNA, Neoplasm; D005557:Formaldehyde