Back to results
US 5,710,000 AGranted
Capturing sequences adjacent to Type-IIs restriction sites for genomic library mapping
AssigneeAffymetrix, Inc.
InventorsRonald J. Sapolsky, Robert J. Lipshutz, Thomas R. Gingeras
Life of this patent
Estimating the term…
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.
Claims11
A method of identifying sequences in a polynucleotide sequence, comprising:
- (a) cleaving the polynucleotide sequence with a first type-IIs endonuclease;
- (b) ligating a first adapter sequence to the polynucleotide sequence cleaved in step (a), said first adapter having a recognition site for a second type-IIs endonuclease, the recognition site being oriented so that the second type-IIs endonuclease has a cleavage site in the polynucleotide sequence;
- (c) cleaving the polynucleotide sequence resulting from step (b), with the second type-IIs endonuclease;
- (d) ligating a second adapter sequence to the polynucleotide sequence cleaved in step (c);
- and
- determining the sequence of nucleotides of the polynucleotide sequence between the first and second adapter sequences.
3 more elements
Dependencies are inferred from the claim text.