Normalization and experimental design for ChIP-chip data

<p>Abstract</p> <p>Background</p> <p>Chromatin immunoprecipitation on tiling arrays (ChIP-chip) has been widely used to investigate the DNA binding sites for a variety of proteins on a genome-wide scale. However, several issues in the processing and analysis of ChIP-chi...

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Main Authors: Alekseyenko Artyom A, Peng Shouyong, Larschan Erica, Kuroda Mitzi I, Park Peter J
Format: Article
Language:English
Published: BMC 2007-06-01
Series:BMC Bioinformatics
Online Access:http://www.biomedcentral.com/1471-2105/8/219
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author Alekseyenko Artyom A
Peng Shouyong
Larschan Erica
Kuroda Mitzi I
Park Peter J
author_facet Alekseyenko Artyom A
Peng Shouyong
Larschan Erica
Kuroda Mitzi I
Park Peter J
author_sort Alekseyenko Artyom A
collection DOAJ
description <p>Abstract</p> <p>Background</p> <p>Chromatin immunoprecipitation on tiling arrays (ChIP-chip) has been widely used to investigate the DNA binding sites for a variety of proteins on a genome-wide scale. However, several issues in the processing and analysis of ChIP-chip data have not been resolved fully, including the effect of background (mock control) subtraction and normalization within and across arrays.</p> <p>Results</p> <p>The binding profiles of <it>Drosophila </it>male-specific lethal (MSL) complex on a tiling array provide a unique opportunity for investigating these topics, as it is known to bind on the X chromosome but not on the autosomes. These large bound and control regions on the same array allow clear evaluation of analytical methods.</p> <p>We introduce a novel normalization scheme specifically designed for ChIP-chip data from dual-channel arrays and demonstrate that this step is critical for correcting systematic dye-bias that may exist in the data. Subtraction of the mock (non-specific antibody or no antibody) control data is generally needed to eliminate the bias, but appropriate normalization obviates the need for mock experiments and increases the correlation among replicates. The idea underlying the normalization can be used subsequently to estimate the background noise level in each array for normalization across arrays. We demonstrate the effectiveness of the methods with the MSL complex binding data and other publicly available data.</p> <p>Conclusion</p> <p>Proper normalization is essential for ChIP-chip experiments. The proposed normalization technique can correct systematic errors and compensate for the lack of mock control data, thus reducing the experimental cost and producing more accurate results.</p>
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spelling doaj.art-406a832cdf1b467a9f4ede333f9f809e2022-12-22T00:08:40ZengBMCBMC Bioinformatics1471-21052007-06-018121910.1186/1471-2105-8-219Normalization and experimental design for ChIP-chip dataAlekseyenko Artyom APeng ShouyongLarschan EricaKuroda Mitzi IPark Peter J<p>Abstract</p> <p>Background</p> <p>Chromatin immunoprecipitation on tiling arrays (ChIP-chip) has been widely used to investigate the DNA binding sites for a variety of proteins on a genome-wide scale. However, several issues in the processing and analysis of ChIP-chip data have not been resolved fully, including the effect of background (mock control) subtraction and normalization within and across arrays.</p> <p>Results</p> <p>The binding profiles of <it>Drosophila </it>male-specific lethal (MSL) complex on a tiling array provide a unique opportunity for investigating these topics, as it is known to bind on the X chromosome but not on the autosomes. These large bound and control regions on the same array allow clear evaluation of analytical methods.</p> <p>We introduce a novel normalization scheme specifically designed for ChIP-chip data from dual-channel arrays and demonstrate that this step is critical for correcting systematic dye-bias that may exist in the data. Subtraction of the mock (non-specific antibody or no antibody) control data is generally needed to eliminate the bias, but appropriate normalization obviates the need for mock experiments and increases the correlation among replicates. The idea underlying the normalization can be used subsequently to estimate the background noise level in each array for normalization across arrays. We demonstrate the effectiveness of the methods with the MSL complex binding data and other publicly available data.</p> <p>Conclusion</p> <p>Proper normalization is essential for ChIP-chip experiments. The proposed normalization technique can correct systematic errors and compensate for the lack of mock control data, thus reducing the experimental cost and producing more accurate results.</p>http://www.biomedcentral.com/1471-2105/8/219
spellingShingle Alekseyenko Artyom A
Peng Shouyong
Larschan Erica
Kuroda Mitzi I
Park Peter J
Normalization and experimental design for ChIP-chip data
BMC Bioinformatics
title Normalization and experimental design for ChIP-chip data
title_full Normalization and experimental design for ChIP-chip data
title_fullStr Normalization and experimental design for ChIP-chip data
title_full_unstemmed Normalization and experimental design for ChIP-chip data
title_short Normalization and experimental design for ChIP-chip data
title_sort normalization and experimental design for chip chip data
url http://www.biomedcentral.com/1471-2105/8/219
work_keys_str_mv AT alekseyenkoartyoma normalizationandexperimentaldesignforchipchipdata
AT pengshouyong normalizationandexperimentaldesignforchipchipdata
AT larschanerica normalizationandexperimentaldesignforchipchipdata
AT kurodamitzii normalizationandexperimentaldesignforchipchipdata
AT parkpeterj normalizationandexperimentaldesignforchipchipdata