Sequencing method for genomic rearrangement detection
AssigneeAgilent Technologies, Inc.
InventorsMargherita Corioni, Douglas N Roberts
Life of this patent
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Abstract
The present disclosure is directed to a single-end sequencing method for improved detection of genomic rearrangements such as deletions, insertions, inversions, and translocations that are present in a polynucleotide. A first priming event allows for sequencing of a target sequence, and a second priming event on an adapter allows for identification of the sequences amplified and tagged by selective amplification. The combination of priming events in the same direction facilitates read alignment and the identification of any genomic rearrangements.
Claims17
A composition for detecting a genomic rearrangement in a target polynucleotide having a first binding site, the composition comprising:
- a target-specific first amplification primer comprising a target-specific primer, a second sequencing priming site or complement thereof, and a target-specific barcode;
- and
- a second amplification primer, wherein the second amplification primer hybridizes to (1) a portion of an adapter attached to the target polynucleotide at a distance from the first priming site, or (2) a second priming site of the target polynucleotide, wherein the second priming site is at a distance from the first priming site;
- wherein the second amplification primer or the adapter comprises a first sequencing priming site or complement thereof, and a barcode;
- and
- wherein amplification of the target polynucleotide with the first and second amplification primers generates polynucleotide amplicons comprising first and second sequencing priming sites for first and second primer extensions in a same direction.
A kit for detecting a genomic rearrangement in a target polynucleotide having a first binding site, the kit comprising:
- a target-specific first amplification primer comprising a target-specific primer, a second sequencing priming site or complement thereof, and a target-specific barcode;
- and
- a second amplification primer, wherein the second amplification primer hybridizes to (1) a portion of an adapter attached to the target polynucleotide at a distance from the first priming site, or (2) a second priming site of the target polynucleotide, wherein the second priming site is at a distance from the first priming site;
- wherein the second amplification primer or the adapter comprises a first sequencing priming site or complement thereof, and a barcode;
- and
- wherein amplification of the target polynucleotide with the first and second amplification primers generates polynucleotide amplicons comprising first and second sequencing priming sites for first and second primer extensions in a same direction.
Dependencies are inferred from the claim text.