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Experimental Annotation of the Human Genome

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This talk will describe the development and application of genomic DNA tiling arrays for the annotation of eukaryotic genomes, with a particular focus on homo sapiens. Tiling arrays are designed to represent all of the DNA comprising a chromosome or other genomic locus, without prior consultation of existing gene annotation. Because they are developed for the unbiased interrogation of genomic sequence, tiling arrays enable the discovery of novel transcripts and regulatory elements in eukaryotic genomes. Two examples of microarray-based genome annotation are presented: large-scale mapping of transcribed sequences and identification of transcription factor-binding sites. Chromosome- and genome-wide transcriptional activity is assessed by probing tiling arrays with normal complex tissue RNA ; the locations of cis-regulatory elements are determined via array hybridisation to chromatin-immunoprecipitated DNA , originally bound in vivo by a transcription factor of interest. These approaches are first demonstrated with amplicon arrays representing all of the non-repetitive DNA of human chromosome 22, then extended to the entire genome using maskless photolithographic DNA synthesis technology. A large-scale tiling array survey revealed the presence of thousands of novel transcribed sequences in addition to known and predicted genes, providing the first global transcription map of the human genome.

This talk is part of the Inference Group series.

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