Universal reagents for rolling circle amplification and methods of use
First Claim
1. A composition comprising a reporter binding agent, an amplification target circle, and DNA polymerase,wherein the reporter binding agent comprises a specific binding molecule and a rolling circle replication primer,wherein the specific binding molecule is specific for a target molecule, wherein the specific binding molecule is not bound to the target molecule, wherein the composition does not comprise tandem sequence DNA, wherein the reporter binding agent, the amplification target circle, and the DNA polymerase are directly associated with each other and form a complex, and wherein the composition is not in an assay or reaction.
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Accused Products
Abstract
Disclosed are compositions and methods useful for labeling and detection of analytes. The compositions generally are associations of three components: reporter binding agents, amplification target circles, and DNA polymerase. The compositions are assembled prior to their use in a rolling circle amplification reaction and can be stored and transported prior to use without substantial loss of activity. The reporter binding agents generally are composed of a specific binding molecule and a rolling circle replication primer. The specific binding molecule can be specific for a target molecule. The rolling circle replication primer has sequence complementary to the amplification target circle. The DNA polymerase can interact with the rolling circle replication primer and amplification target circle. For use as a general reagent, the specific binding molecule is not bound to the target molecule until the composition is used in an assay.
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Citations
66 Claims
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1. A composition comprising a reporter binding agent, an amplification target circle, and DNA polymerase,
wherein the reporter binding agent comprises a specific binding molecule and a rolling circle replication primer, wherein the specific binding molecule is specific for a target molecule, wherein the specific binding molecule is not bound to the target molecule, wherein the composition does not comprise tandem sequence DNA, wherein the reporter binding agent, the amplification target circle, and the DNA polymerase are directly associated with each other and form a complex, and wherein the composition is not in an assay or reaction.
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24. A kit comprising a composition and a buffer for rolling circle amplification,
wherein the composition comprises a reporter binding agent, an amplification target circle, and DNA polymerase, wherein the reporter binding agent comprises a specific binding molecule and a rolling circle replication primer, wherein the specific binding molecule is specific for a target molecule, wherein the specific binding molecule is not bound to the target molecule, wherein the composition does not comprise tandem sequence DNA, wherein the reporter binding agent, the amplification target circle, and the DNA polymerase directly associated with each other and form a complex, wherein the composition is not in an assay or reaction.
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62. A composition comprising a reporter binding agent, an amplification target circle, and DNA polymerase,
wherein the reporter binding agent comprises a specific binding molecule and a rolling circle replication primer, wherein the specific binding molecule is specific for a target molecule, wherein the specific binding molecule is not bound to the target molecule, wherein the composition does not comprise tandem sequence DNA, wherein the reporter binding agent, the amplification target circle, and the DNA polymerase are directly associated with each other and form a complex, wherein the composition is stored at or below about 25° - C. for 1 day or longer prior to use of the composition in an assay or reaction.
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63. A composition comprising a reporter binding agent, an amplification target circle, and DNA polymerase,
wherein the reporter binding agent comprises a specific binding molecule and a rolling circle replication primer, wherein the specific binding molecule is specific for a target molecule, wherein the specific binding molecule is not bound to the target molecule, wherein the composition does not comprise tandem sequence DNA, wherein the reporter binding agent, the amplification target circle, and the DNA polymerase are directly associated with each other and form a complex, wherein the composition is stored at or below about 4° - C. for 10 days or longer prior to use of the composition in an assay or reaction.
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64. A composition comprising a reporter binding agent, an amplification target circle, and DNA polymerase,
wherein the reporter binding agent comprises a specific binding molecule and a rolling circle replication primer, wherein the specific binding molecule is specific for a target molecule, wherein the specific binding molecule is not bound to the target molecule, wherein the composition does not comprise tandem sequence DNA, wherein the reporter binding agent, the amplification target circle, and the DNA polymerase are directly associated with each other and form a complex, wherein the composition is stored at or below about − - 20°
C. for 1 month or longer prior to use of the composition in an assay or reaction.
- 20°
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65. A kit comprising a composition and a buffer for rolling circle amplification,
wherein the composition comprises a reporter binding agent, an amplification target circle, and DNA polymerase, wherein the reporter binding agent comprises a specific binding molecule and a rolling circle replication primer, wherein the specific binding molecule is specific for a target molecule, wherein the specific binding molecule is not bound to the target molecule, wherein the composition does not comprise tandem sequence DNA, wherein the reporter binding agent, the amplification target circle, and the DNA polymerase are directly associated with each other and form a complex, wherein the composition is stored at or below about 25° - C. for 1 day or longer prior to use of the composition in an assay or reaction.
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66. A kit comprising a composition and a buffer for rolling circle amplification,
wherein the composition comprises a reporter binding agent, an amplification target circle, and DNA polymerase, wherein the reporter binding agent comprises a specific binding molecule and a rolling circle replication primer, wherein the specific binding molecule is specific for a target molecule, wherein the specific binding molecule is not bound to the target molecule, wherein the composition does not comprise tandem sequence DNA, wherein the reporter binding agent, the amplification target circle, and the DNA polymerase are directly associated with each and form a complex, wherein the composition is stored at or below about 4° - C. for 10 days or longer prior to use of the composition in an assay or reaction.
Specification