Back to results
US 2009/0023190 A1Published application
Sequence amplification with loopable primers
OwnerKai Qin Lao, Neil A. Strausinventors
Life of this patent
Estimating the term…
Abstract
The present disclosure relates to the amplification of target nucleic acid sequences. This can be accomplished via the use of various primers. The use of these primers, as described herein, results in nucleic acid structures that can reduce the amplification of nonspecific hybridization events (such as primer dimerization) while allowing the amplification of the target nucleic acid sequences.
Claims46
A nucleic acid sequence comprising:
- a 3′ target specific region;
- a first loop forming region;
- a universal region;
- and
- a second loop forming region, wherein said first and second loop forming regions comprise a set of nucleic acid sequences that are configured to hybridize to one another and wherein the universal region is located between the first and second loop forming regions, wherein the universal region is configured so that the nucleic acid sequence can form a self-hybridized structure comprising the universal region, the first loop forming region and the second loop forming region on subsequent amplifications.
2 more elements
A self-hybridizable DNA structure in a PCR amplification mixture, said self-hybridizable DNA sequence comprising:
- a first loop forming region;
- a universal region connected to the first loop forming region;
- a second loop forming region connected to the universal region;
- a first 3′ target specific region connected to the second loop forming region;
- a sequence that is complementary to a second 3′ target specific region, wherein the sequence that is complementary to the second 3′ target specific region is part of a same nucleic acid strand as the first 3′ target specific region;
- a sequence that is complementary to the second loop forming region, wherein the sequence that is complementary to the second loop forming region is connected to the sequence that is complementary to the second 3′ target specific region;
- a sequence that is complementary to the universal region, wherein the a sequence that is complementary to the universal region is connected to the sequence that is complementary to the second loop forming region;
- and
- a sequence that is complementary to the first loop forming region, wherein the sequence that is complementary to the first loop forming region is connected to the sequence that is complementary to the universal region.
6 more elements
A method for nucleic acid amplification, said method comprising:
- allowing a 3′ target specific region of a loopable primer to hybridize to a first part of a target nucleic acid sequence, wherein a loop in the loopable primer is configured to allow a 3′ end of the loopable primer to hybridize to a first part of the target nucleic acid sequence without the remainder of the loopable primer annealing to the target nucleic acid sequence, wherein the loopable primer further comprises:
- a first loop forming region, a universal region, and a second loop forming region, wherein said first and second loop forming regions comprise nucleic acid sequences that hybridize to one another and wherein the universal region is located between the first and second loop forming regions;
- extending the loopable primer that is hybridized to the target nucleic acid sequence to form an extended loopable primer;
- allowing an additional loopable primer to hybridize to a complementary part of the target nucleic acid sequence on the extended loopable primer;
- and
- extending the loopable primer to form a double-extended loopable primer.
3 more elements
A nucleic acid sequence comprising:
- a 3′ target specific region;
- a first loop forming region;
- a universal region;
- a noncomplementary region;
- and
- a second loop forming region, wherein said first and second loop forming regions comprise a set of nucleic acid sequences that are configured to hybridize to one another and wherein the universal region is located between the first and second loop forming regions, wherein the noncomplementary region is located between the first and second loop forming regions, wherein the universal region is configured so that a sequence associated with a target sequence that has been amplified by the nucleic acid sequence can form a self-hybridized structure comprising the universal region, the noncomplementary region, the first loop forming region, and the second loop forming region on subsequent amplifications, wherein said nucleic acid is created by deliberately selecting the sequence of the first loop forming region so that it hybridizes to the second loop forming region, and wherein said nucleic acid is created by deliberately selecting a universal region.
3 more elements
Dependencies are inferred from the claim text.