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Ruler Arrays Reveal Haploid Genomic Structural Variation

ABSTRACT: A ruler array combines novel sample preparation protocols, tiling genomic microarrays, and a new computational method to turn each probe on a microarray into a ruler that measures the physical distance between defined genomic sequences regardless of the intervening sequence. The ruler array protocol involves (1) genomic DNA purification (2) digestion with a restriction enzyme (3) ligation of a biotinylated adapter molecule to the cut sites (4) purification on streptavidin beads (5) polymerase extensions from a primer complementary to the adapter using labeled dNTPs and (6) hybridization to a tiling microarray

ORGANISM(S): Saccharomyces cerevisiae

PROVIDER: GSE23524 | GEO | 2010/09/16

SECONDARY ACCESSION(S): PRJNA131777

REPOSITORIES: GEO

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Ruler Arrays Reveal Haploid Genomic Structural Variation

Project description:A ruler array combines novel sample preparation protocols, tiling genomic microarrays, and a new computational method to turn each probe on a microarray into a ruler that measures the physical distance between defined genomic sequences regardless of the intervening sequence. The ruler array protocol involves (1) genomic DNA purification (2) digestion with a restriction enzyme (3) ligation of a biotinylated adapter molecule to the cut sites (4) purification on streptavidin beads (5) polymerase extensions from a primer complementary to the adapter using labeled dNTPs and (6) hybridization to a tiling microarray Two replicates of our ruler array experiment to detect insertions and deletions between two genomic samples of strains FY4 and Sigma1278b.

2010-09-16 | E-GEOD-23524 | biostudies-arrayexpress

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Ruler elements in chromatin remodelers set nucleosome array spacing and phasing

Project description:Arrays of regularly spaced nucleosomes dominate chromatin and are often phased, i.e., aligned at reference sites like active promoters. How distances between nucleosomes and distances between phasing sites and nucleosomes are determined remained unclear, specifically, the role of ATP dependent chromatin remodelers in it. Here, we used a genome-wide reconstitution system to probe how yeast remodelers generate phased nucleosome arrays. We find that remodelers bear a structural element named the ‘ruler’ that sets nucleosome spacing, in the order Chd1 < ISW1a < ISW2 < INO80. Structure-based mutagenesis confirmed the functional significance of the ruler element in INO80. Differences in the ruler elements of different remodelers explain the observed nucleosome array features. More generally, we propose that remodelers use their rulers to regulate the direction of nucleosome sliding in response to nucleosome density and environment, leading to nucleosome positioning relative to other nucleosomes, DNA bound factors or DNA sequence elements.

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Project description:DNA samples of haploid ES cells and control DNA were compared to genomic DNA of the C57B/6 inbred mouse strain on a NimbleGen Mouse CGH 3x720K Whole-Genome Tiling Array (Build MM9), (GPL10989) DNA samples of haploid ES cells and control DNA were compared to genomic DNA of the 129/Sv inbred mouse strain on a NimbleGen Mouse CGH 3x720K Whole-Genome Tiling Array (Build MM9), (GPL10989)

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