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US 2021/0172030 A1Published application
Method for detecting a tandem repeat
AssigneeImperial College of Science, Technology and Medicine
InventorsPantelis GEORGIOU, Ling-Shan YU, Kenny Malpartida CARDENAS, Matthew C. FISHER, Jesus Rodriguez MANZANO, Nicolas MOSER
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Abstract
The present application relates to methods for detecting a tandem repeat in a nucleic acid sequence under isothermal conditions using primers.
Claims25
A method for detecting a first allele of a single nucleotide polymorphism (SNP) in a nucleic acid sequence, wherein the method comprises:
- (a) amplifying under isothermal conditions and stringent conditions a nucleic acid sequence from a sample, in a reaction mixture comprising
- (i) the nucleic acid sequence,
- (ii) a nucleic acid polymerase,
- (iii) a nucleoside triphosphate mixture,
- (iv) a forward inner primer targeting the first allele (FIPa1), comprising a F1c region which anneals to a F1 region of the nucleic acid sequence in the presence of the first allele and a F2 region which anneals to a F2c region of the nucleic acid sequence,
- (v) a backward inner primer targeting the first allele (BIPa1), comprising a B1c region which anneals to a B1 region of the nucleic acid sequence in the presence of the first allele and a B2 region which anneals to a B2c region of the nucleic acid sequence,
- (vi) a forward blocking primer targeting a second allele of the SNP (FBa2), comprising a B1c region which anneals to a B1 region of the nucleic acid sequence in the presence of the second allele, and
- (vii) a backward blocking primer targeting the second allele (BBa2), comprising a F1c region which anneals to a F1 region of the nucleic acid sequence in the presence of the second allele,
- wherein the F1 region of the nucleic acid sequence and the B1 region of the nucleic acid sequence both contain the SNP, and
- (b) detecting an amplified nucleic acid sequence, wherein the detection of the amplified nucleic acid sequence indicates the presence of the first allele of the SNP.
7 more elements
A kit comprising:
- (i) a forward inner primer targeting a first allele (FIPa1), comprising a F1c region which anneals to a F1 region of a nucleic acid sequence in the presence of the first allele of a single nucleotide polymorphism (SNP) in a nucleic acid sequence and a F2 region which anneals to a F2c region of the nucleic acid sequence,
- (ii) a backward inner primer targeting the first allele (B1Pa1), comprising a B1c region which anneals to a B1 region of the nucleic acid sequence in the presence of the first allele and a B2 region which anneals to a B2c region of the nucleic acid sequence,
- (iii) a forward blocking primer targeting a second allele of the SNP (FBa2), comprising a B1c region which anneals to a B1 region of the nucleic acid sequence in the presence of the second allele, and
- (iv) a backward blocking primer targeting the second allele (BBa2), comprising a F1c region which anneals to a F1 region of the nucleic acid sequence in the presence of the second allele,
- wherein the F1 region of the nucleic acid sequence and the B1 region of the nucleic acid sequence both contain the SNP.
2 more elements
Dependencies are inferred from the claim text.