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Old 06-08-2011, 03:50 AM   #1
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Default ChIP-Seq: Systematic bias in high-throughput sequencing data and its correction by BE

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Systematic bias in high-throughput sequencing data and its correction by BEADS.

Nucleic Acids Res. 2011 Jun 6;

Authors: Cheung MS, Down TA, Latorre I, Ahringer J

Genomic sequences obtained through high-throughput sequencing are not uniformly distributed across the genome. For example, sequencing data of total genomic DNA show significant, yet unexpected enrichments on promoters and exons. This systematic bias is a particular problem for techniques such as chromatin immunoprecipitation, where the signal for a target factor is plotted across genomic features. We have focused on data obtained from Illumina's Genome Analyser platform, where at least three factors contribute to sequence bias: GC content, mappability of sequencing reads, and regional biases that might be generated by local structure. We show that relying on input control as a normalizer is not generally appropriate due to sample to sample variation in bias. To correct sequence bias, we present BEADS (bias elimination algorithm for deep sequencing), a simple three-step normalization scheme that successfully unmasks real binding patterns in ChIP-seq data. We suggest that this procedure be done routinely prior to data interpretation and downstream analyses.

PMID: 21646344 [PubMed - as supplied by publisher]



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