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The T7-primer is a source of experimental bias and introduces variability between microarray platforms.

Authors :
Ron M Kerkhoven
Daoud Sie
Marja Nieuwland
Mike Heimerikx
Jorma De Ronde
Wim Brugman
Arno Velds
Source :
PLoS ONE, Vol 3, Iss 4, p e1980 (2008)
Publication Year :
2008
Publisher :
Public Library of Science (PLoS), 2008.

Abstract

Eberwine(-like) amplification of mRNA adds distinct 6-10 bp nucleotide stretches to the 5' end of amplified RNA transcripts. Analysis of over six thousand microarrays reveals that probes containing motifs complementary to these stretches are associated with aberrantly high signals up to a hundred fold the signal observed in unaffected probes. This is not observed when total RNA is used as target source. Different T7 primer sequences are used in different laboratories and platforms and consequently different T7 primer bias is observed in different datasets. This will hamper efforts to compare data sets across platforms.

Subjects

Subjects :
Medicine
Science

Details

Language :
English
ISSN :
19326203
Volume :
3
Issue :
4
Database :
Directory of Open Access Journals
Journal :
PLoS ONE
Publication Type :
Academic Journal
Accession number :
edsdoj.58d316ba760460dbc22fd6a6febd9ed
Document Type :
article
Full Text :
https://doi.org/10.1371/journal.pone.0001980