Hybridization buffer systems impact the quality of filter array data.

Abstract:

INTRODUCTION:cDNA microarray technology has greatly facilitated mechanistic studies in pharmacology and toxicology. A clean hybridization with minimal background is critical for successful microarray analysis and is highly desired. However, clean hybridization alone is not enough; verification is needed. METHODS:Total RNA was isolated from the livers of acetaminophen-intoxicated mice and was subjected to cDNA microarray analyses using ExpressHyb, ULTRArrayHyb or MicroHyb on nylon membranes. Real-time RT-PCR analyses were performed for verification. RESULTS:We have demonstrated in this paper that hybridization systems can significantly impact the quality of array data. MicroHyb produced very clean hybridizations, but some results could not be confirmed by real-time RT-PCR and in accord with biological responses. The hybridization images from ExpressHyb were not always clean, but were reliable. The sensitivity of ULTRArrayHyb was moderate. CONCLUSION:This study has indicated the importance of selecting hybridization buffers in membrane arrays and recommended real-time RT-PCR for follow-up analysis. Gene expression changes should also be correlated with biological significance.

authors

Liu J,Walker N,Waalkes MP

doi

10.1016/j.vascn.2004.02.001

keywords:

subject

Has Abstract

pub_date

2004-07-01 00:00:00

pages

67-71

issue

1

eissn

1056-8719

issn

1873-488X

pii

S1056-8719(04)00006-1

journal_volume

50

pub_type

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