Project description:We characterised the genome organization in non-bilaterian animals (sponge Ephydatia muelleri, ctenophores Mnemiopsis leidyi and Hormiphora californensis, placozoans Trichoplax adhaerens and Cladtertia collaboinventa, and cnidarian Nematostella vectensis) and close unicellular relatives (ichthyosporeans Sphaeroforma arctica, filasterean Capsaspora owczarzaki and choanoflagellate Salpingoeca rosetta) by combining high-resolution chromosome conformation capture (Micro-C) with profiling of chromatin marks (ChIP-seq), accessible genomic regions (ATAC-seq) and gene expression (MARS-seq).
Project description:We characterised the genome organization in non-bilaterian animals (sponge Ephydatia muelleri, ctenophores Mnemiopsis leidyi and Hormiphora californensis, placozoans Trichoplax adhaerens and Cladtertia collaboinventa, and cnidarian Nematostella vectensis) and close unicellular relatives (ichthyosporeans Sphaeroforma arctica, filasterean Capsaspora owczarzaki and choanoflagellate Salpingoeca rosetta) by combining high-resolution chromosome conformation capture (Micro-C) with profiling of chromatin marks (ChIP-seq), accessible genomic regions (ATAC-seq) and gene expression (MARS-seq).
Project description:Microarrays are useful tools for detecting and quantifying specific functional and phylogenetic genes in natural microbial communities. In order to track uncultivated microbial genotypes and their close relatives in an environmental context, we designed and implemented a “genome proxy” microarray that targets microbial genome fragments recovered directly from the environment. Fragments consisted of sequenced clones from large-insert genomic libraries from microbial communities in Monterey Bay, the Hawaii Ocean Time-series station ALOHA, and Antarctic coastal waters. In a prototype array, we designed probe sets to thirteen of the sequenced genome fragments and to genomic regions of the cultivated cyanobacterium Prochlorococcus MED4. Each probe set consisted of multiple 70-mers, each targeting an individual ORF, and distributed along each ~40-160kbp contiguous genomic region. The targeted organisms or clones, and close relatives, were hybridized to the array both as pure DNA mixtures and as additions of cells to a background of coastal seawater. This prototype array correctly identified the presence or absence of the target organisms and their relatives in laboratory mixes, with negligible cross-hybridization to organisms having ≤~75% genomic identity. In addition, the array correctly identified target cells added to a background of environmental DNA, with a limit of detection of ~0.1% of the community, corresponding to ~10^3 cells/ml in these samples. Signal correlated to cell concentration with an R2 of 1.0 across six orders of magnitude. In addition the array could track a related strain (at 86% genomic identity to that targeted) with a linearity of R2=0.9999 and a limit of detection of ~1% of the community. Closely related genotypes were distinguishable by differing hybridization patterns across each probe set. This array’s multiple-probe, “genome-proxy” approach and consequent ability to track both target genotypes and their close relatives is important for the array’s environmental application given the recent discoveries of considerable intra-population diversity within marine microbial communities. Keywords: target addition experiment, proof-of-concept for GPL6012
2007-10-31 | GSE9384 | GEO
Project description:Development and characterization of microsatellite markers for Antennaria corymbosa (Asteraceae) and close relatives
Project description:We characterised the genome organization in non-bilaterian animals (sponge Ephydatia muelleri, ctenophores Mnemiopsis leidyi and Hormiphora californensis, placozoans Trichoplax adhaerens and Cladtertia collaboinventa, and cnidarian Nematostella vectensis) and close unicellular relatives (ichthyosporeans Sphaeroforma arctica, filasterean Capsaspora owczarzaki and choanoflagellate Salpingoeca rosetta) by combining high-resolution chromosome conformation capture (Micro-C) with profiling of chromatin marks (ChIP-seq), accessible genomic regions (ATAC-seq) and gene expression (MARS-seq).