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Snijders Scientific a tiling resolution dna microarray with complete coverage of the human genome
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mV H –CTCF consensus motif is a major determinant of CTCF binding at the murine IgH locus. ChIP with an α-CTCF antibody was performed on CD19 + pro-B cells that were isolated from 8- to 9-week-old RAG2 −/− mice and expanded for 3 days in the presence of IL-7 (10 ng/ml). Cy3-labeled input DNA and Cy5-labeled immunoprecipitated DNA were hybridized to customized tiling DNA microarrays, and peaks were called by the Ringo method . a , area-proportional Venn diagram showing the overlap between predicted CTCF-binding sites in the V H domain of the murine IgH locus ( cyan ) and observed CTCF peaks ( yellow ). b , area-proportional Venn diagram showing the overlap between predicted CTCF-binding sites ( cyan ); CTCF peaks were observed by ChIP-chip ( yellow ), and CTCF peaks were observed previously by ChIP-seq . c , enoLOGOS representation of three distinct sequence motifs identified by MEME analysis of the 79 CTCF peaks observed by ChIP-chip that did not contain an mV H –CTCF consensus motif.

Journal: The Journal of Biological Chemistry

Article Title: The murine IgH locus contains a distinct DNA sequence motif for the chromatin regulatory factor CTCF

doi: 10.1074/jbc.RA118.007348

Figure Lengend Snippet: mV H –CTCF consensus motif is a major determinant of CTCF binding at the murine IgH locus. ChIP with an α-CTCF antibody was performed on CD19 + pro-B cells that were isolated from 8- to 9-week-old RAG2 −/− mice and expanded for 3 days in the presence of IL-7 (10 ng/ml). Cy3-labeled input DNA and Cy5-labeled immunoprecipitated DNA were hybridized to customized tiling DNA microarrays, and peaks were called by the Ringo method . a , area-proportional Venn diagram showing the overlap between predicted CTCF-binding sites in the V H domain of the murine IgH locus ( cyan ) and observed CTCF peaks ( yellow ). b , area-proportional Venn diagram showing the overlap between predicted CTCF-binding sites ( cyan ); CTCF peaks were observed by ChIP-chip ( yellow ), and CTCF peaks were observed previously by ChIP-seq . c , enoLOGOS representation of three distinct sequence motifs identified by MEME analysis of the 79 CTCF peaks observed by ChIP-chip that did not contain an mV H –CTCF consensus motif.

Article Snippet: Tiling genomic DNA microarrays were custom-designed (NimbleGen Systems, Inc.) based on the mm9 release of the IgH locus sequence (murine chr12: 114,341,024–117,349,200).

Techniques: Binding Assay, Isolation, Labeling, Immunoprecipitation, ChIP-chip, ChIP-sequencing, Sequencing