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research article

Absolute mRNA concentrations from sequence-specific calibration of oligonucleotide arrays

Hekstra, Doeke
•
Taussig, Alexander R.
•
Magnasco, Marcelo
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2003
Nucleic Acids Research

Oligonucleotide microarrays are based on the hybridization of labeled mRNA molecules to short length oligonucleotide probes on a glass surface. Two effects have been shown to affect the raw data: the sequence dependence of the probe hybridization properties and the chemical saturation resulting from surface adsorption processes. We address both issues simultaneously using a physically motivated hybridization model. Based on publicly available calibration data sets, we show that Langmuir adsorption accurately describes GeneChip hybridization, with model parameters that we predict from the sequence composition of the probes. Because these parameters have physical units, we are able to estimate absolute mRNA concentrations in picomolar. Additionally, by accounting for chemical saturation, we substantially reduce the compressive bias of differential expression estimates that normally occurs toward high concentrations.

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Type
research article
DOI
10.1093/nar/gkg283
Author(s)
Hekstra, Doeke
Taussig, Alexander R.
Magnasco, Marcelo
Naef, Felix  
Date Issued

2003

Publisher

Oxford University Press

Published in
Nucleic Acids Research
Volume

31

Issue

7

Start page

1962

End page

8

Editorial or Peer reviewed

REVIEWED

Written at

OTHER

EPFL units
UPNAE  
Available on Infoscience
November 1, 2010
Use this identifier to reference this record
https://infoscience.epfl.ch/handle/20.500.14299/56506
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