Filtered shotgun sequencing of complex eukaryotic genomes
AssigneeCold Spring Harbor Laboratory
InventorsW. Richard McCombie, Robert A. Martienssen
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Abstract
This invention provides methods by which repetitive elements can be selectively removed from genomic libraries made from complex eukaryotic genomes. In particular, the invention relates to affecting the efficiency of recovery of novel genes and regulatory sequences, by use of methylation restrictive bacteria, cell-free enzymes, and methyl-binding proteins.
Claims45
A genomic cloning method for identifying DNA segments containing genes in complex genomes, said method comprising:
- constructing a genomic library in an environment that eliminates repetitive sequences, said library comprising fragments of genomic DNA;
- inserting said genomic DNA into a suitable vector, and
- characterizing said DNA segment. 2.
A genomic cloning method for identifying DNA segments containing genes in complex genomes, said method comprising:
- constructing a genomic library in an environment that eliminates repetitive sequences, said library comprising fragments of genomic DNA;
- said environment established using mechanical shearing of said genomic DNA, selective hybridization at Cot values of between approximately 0.01 and 100, followed by hydroxylapatite chromatography;
- inserting said genomic DNA into a suitable vector, and
- characterizing said DNA segment.
A genomic cloning method for identifying DNA segments containing genes in complex genomes, said method comprising:
- constructing a genomic library in a methylation restrictive environment, said library comprising fragments of genomic DNA;
- inserting said genomic DNA into a suitable vector, and
- characterizing said DNA segment.
A genomic cloning-method for identifying DNA segments containing genes in complex genomes, said method comprising:
- constructing a genomic library in an environment that eliminates repetitive sequences by selective protein binding of said genomic DNA, said library comprising fragments of genomic DNA;
- inserting said genomic DNA into a suitable vector, and
- characterizing said DNA segment.
Dependencies are inferred from the claim text.