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US 12,703,880 B2Granted
Method for asymmetric amplification of multiple target nucleic acids
AssigneeXiamen University
InventorsQiuying Huang, Qingge Li
Life of this patent
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Abstract
Provided is multiplex and asymmetric amplification of nucleic acid molecules. In particular, provided is a method for simultaneous and asymmetric amplification of one or more target nucleic acids in a sample. The method can simultaneously and asymmetrically amplify multiple target nucleic acids existing in a sample, and can simultaneously produce large number of single stranded products.
Claims19
A method for amplifying one or more target nucleic acids in a sample, comprising:
- (1) providing:
- (i) a sample containing one or more target nucleic acids;
- (ii) a first universal primer and a second universal primer;
- and (iii) a target-specific primer pair for each target nucleic acid to be amplified;
- wherein,
- the first universal primer comprises a first universal sequence;
- the second universal primer comprises a second universal sequence, wherein the second universal sequence comprises the first universal sequence and additionally comprises at least one nucleotide at a 3′ end of the first universal sequence;
- the target-specific primer pair is capable of amplifying the target nucleic acid and comprises a forward primer and a reverse primer, wherein the forward primer comprises the first universal sequence and a forward nucleotide sequence specific to the target nucleic acid, and the forward nucleotide sequence is located at the 3′ end of the first universal sequence;
- the reverse primer comprises the second universal sequence and a reverse nucleotide sequence specific to the target nucleic acid, and the reverse nucleotide sequence is located at a 3′ end of the second universal sequence;
- and, the second universal sequence is not completely complementary to a complementary sequence of the forward primer;
- and
- (2) amplifying the target nucleic acids in the sample through a PCR reaction using the first universal primer, the second universal primer and the target-specific primer pair under a condition that allows nucleic acid amplification.
9 more elements
A primer set, which comprises:
- a first universal primer and a second universal primer, and, one or more target-specific primer pairs;
- wherein,
- the first universal primer comprises a first universal sequence;
- the second universal primer comprises a second universal sequence, wherein the second universal sequence comprises the first universal sequence and additionally comprises at least one nucleotide at a 3′ end of the first universal sequence;
- each target-specific primer pair is capable of amplifying a target nucleic acid and comprises a forward primer and a reverse primer, wherein the forward primer comprises the first universal sequence and a forward nucleotide sequence specific to the target nucleic acid, and the forward nucleotide sequence is located at the 3′ end of the first universal sequence;
- the reverse primer comprises the second universal sequence and a reverse nucleotide sequence specific to the target nucleic acid, and the reverse nucleotide sequence is located at a 3′ end of the second universal sequence;
- and, the second universal sequence is not completely complementary to a complementary sequence of the forward primer.
4 more elements
Dependencies are inferred from the claim text.