Document Detail


Providing a stable methodological basis for comparing transcript abundance of developing embryos using microarrays.
MedLine Citation:
PMID:  20479066     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
High throughput methods deliver large amount of data serving to describe the physiological treatment that is being studied. In the case of microarrays, there would be a clear benefit to integrate the published data sets. However, the numerous methodological discrepancies between microarray platforms make this comparison impossible. This incompatibility is magnified when considering the peculiar context of transcript management in early embryogenesis. The total RNA content is known to profoundly fluctuate during development. In addition, the mRNA population is subjected to poly(A) tail shortening and elongating events, a characteristic of stored and recruited messengers. These intrinsic factors need to be considered when interpreting any transcript abundance profiles during early development. As a consequence, many methodological details affect microarray platform performances and prevent compatibility. In an effort to maximize our microarray platform performance, we determined the various sources of variation for every one of the main steps leading to the production of microarray data. The five main steps involved in sample preparation were evaluated, as well as conditions for post-hybridization validation by qRT-PCR. These determinations were essential for the implementation of standardized procedures for our Research Network but they can also provide insight into the compatibility issues that the microarray community is now facing.
Authors:
Isabelle Gilbert; Sara Scantland; Eve-Lyne Sylvestre; Isabelle Dufort; Marc-André Sirard; Claude Robert
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Publication Detail:
Type:  Comparative Study; Journal Article; Research Support, Non-U.S. Gov't     Date:  2010-05-17
Journal Detail:
Title:  Molecular human reproduction     Volume:  16     ISSN:  1460-2407     ISO Abbreviation:  Mol. Hum. Reprod.     Publication Date:  2010 Aug 
Date Detail:
Created Date:  2010-07-21     Completed Date:  2010-12-22     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9513710     Medline TA:  Mol Hum Reprod     Country:  England    
Other Details:
Languages:  eng     Pagination:  601-16     Citation Subset:  IM    
Affiliation:
Centre de recherche en biologie de la reproduction, Département des Sciences animales, Université Laval, Québec, Canada.
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MeSH Terms
Descriptor/Qualifier:
Embryonic Development*
Humans
Microarray Analysis / methods*
RNA / biosynthesis
Reproductive Techniques, Assisted*
Reverse Transcriptase Polymerase Chain Reaction
Transcription, Genetic
Chemical
Reg. No./Substance:
63231-63-0/RNA

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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