Methods and reagents for amplifying nucleic acids
AssigneeThe United States of America, as represented by the Secretary, Department of Health and Human Services
InventorsJothikumar Narayanan, Prithiviraj Jothikumar, Vincent Hill
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Abstract
This disclosure related to methods and reagents for isothermal amplification of nucleic acid molecules. In some embodiments, methods are provided for amplification of a nucleic acid molecule from a biological sample. Additional embodiments include identification of a target nucleic acid molecule in a biological sample using the disclosed amplification methods.
Claims29
A method of amplifying a target nucleic acid molecule comprising from 5′ to 3′, a 5′-end region, a center region, and a 3′-end region, wherein each region is non-overlapping and comprises at least 10 consecutive nucleotides, comprising:
- contacting the target nucleic acid molecule under conditions sufficient for isothermal amplification with an effective amount of:
- a forward turn-back primer comprising a first segment comprising at least 10 consecutive nucleotides capable of hybridizing to the center region and a second segment comprising at least 10 consecutive nucleotides capable of hybridizing to the complement of the 5′-end region, wherein the first segment is 5′ to the second segment;
- and
- a reverse turn-back primer comprising a first segment comprising at least 10 consecutive nucleotides capable of hybridizing to the complement of the center region and a second segment comprising at least 10 consecutive nucleotides capable of hybridizing to the 3′-end region, wherein the first segment is 5′ to the second segment;
- and
- amplifying the target nucleic acid molecule using isothermal amplification.
Dependencies are inferred from the claim text.