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Stan/Yale Mouse TFBS

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Stan/Yale Mouse TFBS Documentation

Description

This track shows probable binding sites of the specified transcription factors (TFs)in the given cell types as determined by chromatin immunoprecipitation followed by high throughput sequencing (ChIP-Seq). Each experiment is associated with an input signal, which represents the control condition where immunoprecipitation with non-specific immunoglobulin was performed in the same cell type. For each experiment (cell type vs. antibody) this track shows a graph of enrichment for TF binding (Signal), along with sites that have the greatest evidence of transcription factor binding, as identified by the PeakSeq algorithm (Peaks).

The sequence reads, quality scores, and alignment coordinates from these experiments are available for download.

Methods

Cell growth protocols and instructions for obtaining cells can be found at XXXXX. For details on the chromatin immunoprecipitation protocol used, see Euskirchen et. al. (2007), Rozowsky et. al. (2009) and Auerbach et. al. (2009).

DNA recovered from the precipitated chromatin was sequenced on the Illumina (Solexa) sequencing platform and mapped to the genome using the Eland alignment program. ChIP-seq data was scored based on sequence reads (length ~30 bps) that align uniquely to the human genome. From the mapped tags a signal map of ChIP DNA fragments (average fragment length ~ 200 bp) was constructed where the signal height is the number of overlapping fragments at each nucleotide position in the genome.

For each 1 Mb segment of each chromosome a peak height threshold was determined by requiring a false discovery rate <= 0.01 when comparing the number of peaks above threshold as compared the number obtained from multiple simulations of a random null background with the same number of mapped reads (also accounting for the fraction of mapable bases for sequence tags in that 1 Mb segment). The number of mapped tags in a putative binding region is compared to the normalized (normalized by correlating tag counts in genomic 10 kb windows) number of mapped tags in the same region from an input DNA control. Using a binomial test, only regions that have a p-value <= 0.01 are considered to be significantly enriched compared to the input DNA control.

Expression data supporting the TFBS data can be found in the Stan/Yale Poly-A tracks (coming soon).


Credits

These data were generated and analyzed by the labs of Michael Snyder [http://snyderlab.stanford.edu/ ] at Stanford University and Sherman Weissman [1] at Yale University Contact: pcayting@stanford.edu.


References

Auerbach RK, Euskirchen G, Rozowsky J, Lamarre-Vincent N, Moqtaderi Z, Lefrançois P, Struhl K, Gerstein M, Snyder M. Mapping accessible chromatin regions using Sono-Seq. Proc Natl Acad Sci U S A. 2009 Sep 1;106(35):14926-31. [2]

Rozowsky J, Euskirchen G, Auerbach RK, Zhang ZD, Gibson T, Bjornson R, Carriero N, Snyder M, Gerstein MB. PeakSeq enables systematic scoring of ChIP-seq experiments relative to controls. Nature Biotech. 2009;27:66-75 [3]

Robertson G, Hirst M, Bainbridge M, Bilenky M, Zhao Y, Zeng T, Euskirchen G, Bernier B, Varhol R, Delaney A et al. Genome-wide profiles of STAT1 DNA association using chromatin immunoprecipitation and massively parallel sequencing. Nat. Methods 2007;4:651-7. [4]

Euskirchen GM, Rozowsky JS, Wei C, Lee WH, Zhang ZD, Hartman S, Emanuelsson O, Stolc V, Weissman S, Gerstein MB et al. Mapping of transcription factor binding regions in mammalian cells by ChIP: comparison of array- and sequencing-based technologies. Genome Res. 2007;17:898-909. [5]

Martone R, Euskirchen G, Bertone P, Hartman S, Royce TE, Luscombe NM, Rinn JL, Nelson FK, Miller P, Gerstein M et al. Distribution of NF-kappaB-binding sites across human chromosome 22. Proc Natl Acad Sci U S A. 2003 Oct;100(21):12247-52.[6]