Adaptors for nucleic acid constructs in transmembrane sequencing
AssigneeOxford Nanopore Technologies PLC
InventorBrian McKeown
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Abstract
The invention relates to adaptors for sequencing nucleic acids. The adaptors may be used to generate single stranded constructs of nucleic acid for sequencing purposes. Such constructs may contain both strands from a double stranded deoxyribonucleic acid (DNA) or ribonucleic acid (RNA) template. The invention also relates to the constructs generated using the adaptors, methods of making the adaptors and constructs, as well as methods of sequencing double stranded nucleic acids.
Claims11
-34. (canceled)
A method of identifying the source of origin of a template DNA comprising:
- (i) providing a first sample of template DNA from a first source;
- (ii) ligating a first hairpin adaptor to the template DNA provided in (i), wherein the first hairpin adaptor comprises (a) a double stranded DNA region formed by intramolecular hybridisation between complementary 5′ and 3′ terminal nucleotides of a single stranded DNA oligonucleotide and (b) a single stranded hairpin loop comprising a first nucleic acid sequence identifier;
- (iii) providing a second sample of template DNA from a second source;
- (iv) ligating a second hairpin adaptor to the template DNA provided in (iii), wherein the second hairpin adaptor comprises (a) a double stranded DNA region formed by intramolecular hybridisation between complementary 5′ and 3′ terminal nucleotides of a single stranded DNA oligonucleotide and (b) a single stranded hairpin loop comprising a second nucleic acid sequence identifier;
- (v) mixing the first and second samples of ligated template DNA produced in steps (ii) and (iv);
- (vi) sequencing the ligated template DNA mixture to determine the identity of the nucleic acid sequence identifiers;
- (vii) identifying the source of origin of the template DNA from the identified nucleic acid sequence identifiers.
A pair of adaptors for sequencing nucleic acids, each first and second adaptor of the adaptor pair comprising;
- (i) a double stranded DNA region formed by intramolecular hybridisation between complementary 5′ and 3′ terminal nucleotides of a single stranded DNA oligonucleotide, and
- (ii) a single stranded hairpin loop,
- wherein the single stranded hairpin loop of each of the first and second adaptor pair comprises a different nucleic acid sequence identifier to identify nucleic acids originating from separate individual sources.
Dependencies are inferred from the claim text.