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US 2025/0283152 A1Published application
Nucleic acid amplification using promoter primers
AssigneeSherlock Biosciences, Inc.
InventorsJonathan S. Gootenberg, Omar O. Abudayyeh, Xiang Li, Mary Wilson, William Jeremy Blake
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Abstract
The present disclosure provides methods and compositions for nucleic acid amplification.
Claims34
A method comprising:
- i. contacting a templated nucleic acid synthesis target with:
- (a) a plurality of primers, wherein:
- (1) the plurality includes at least one set of forward and reverse primers, each of which includes a templated nucleic acid synthesis target hybridization element selected so that the primers of the pair bind to opposite strands of a templated nucleic acid synthesis target, flanking a target sequence of interest;
- (2) the plurality of primers furthermore includes at least two T7 promoter sequence elements;
- and
- (b) amplification reagents;
- ii. incubating the templated nucleic acid synthesis target, plurality of primers;
- and
- amplification reagents so an amplified nucleic acid comprising the target sequence of interest and the at least two T7 promoter sequence elements is generated;
- iii. contacting the amplified nucleic acid comprising the target sequence of interest with a CRISPR-Cas detection composition;
- and
- iv. detecting the amplified nucleic acid.
9 more elements
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A composition comprising:
- (a) a plurality of primers, wherein the plurality includes at least one set of forward and reverse primers, each of which includes a templated nucleic acid synthesis target hybridization element selected so that at least one forward primer and at least one reverse primer bind to opposite strands of a template nucleic acid synthesis target, flanking a target sequence of interest;
- wherein the plurality of primers furthermore includes at least two T7 promoter sequence elements and each templated nucleic acid synthesis target hybridization element is at least 80% complementary to its hybridization site in the template nucleic acid synthesis target;
- (b) amplification reagents;
- and
- (c) a CRISPR-Cas detection composition.
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Dependencies are inferred from the claim text.