Capturing sequences adjacent to type-IIs restriction sites for genomic library mapping
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Abstract
The present invention relates to novel methods for sequencing and mapping genetic markers in polynucleotide sequences using Type-IIs restriction endonucleases. The methods herein described result in the “capturing” and determination of specific oligonucleotide sequences located adjacent to Type-IIs restriction sites. The resulting sequences are useful as effective markers for use in genetic mapping, screening and manipulation.
73 Citations
17 Claims
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1-12. -12. (canceled)
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13. A composition comprising at least two oligonucleotide probes, which specifically hybridize to a polymorphism in a DNA sample, and an amplified mixture of DNA isolated from a genome, wherein the amplified mixture of DNA is made by cleaving a genomic DNA sample with at least one restriction enzyme, thereby providing restriction fragments;
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ligating adapter nucleic acids to the DNA restriction fragments;
providing primers that are complementary to the adapter nucleic acids; and
,amplifying the DNA restriction fragments by the polymerase chain reaction by extending the primers, thereby providing the amplified mixture of DNA. - View Dependent Claims (14)
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15. A method of characterizing a nucleic acid, comprising:
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providing at least one oligonucleotide probe which specifically hybridizes to a polymorphic genetic linkage marker in a genomic DNA sample;
amplifying a mixture of nucleic acids comprising a group of genome fragments comprising polymorphisms, thereby providing an amplified nucleic acid mixture of genome fragments; and
hybridizing the at least one oligonucleotide probe to the amplified nucleic acid mixture, thereby detecting at least one nucleic acid fragment in said amplified mixture. - View Dependent Claims (16, 17)
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Specification