Doyle, Charles M.Jen, George C.Pennie & Edmonds LLP
인용정보
피인용 횟수 :
21인용 특허 :
4
초록▼
The present invention provides reversibly blocked primers for use in the amplification of a nucleic acid sequence. Amplifications carried out using the blocked primers result in less non-specific amplification product, in particular, primer dimer, and a concomitant greater yield of the intended ampl
The present invention provides reversibly blocked primers for use in the amplification of a nucleic acid sequence. Amplifications carried out using the blocked primers result in less non-specific amplification product, in particular, primer dimer, and a concomitant greater yield of the intended amplification product compared to amplifications carried out using unblocked primers.
대표청구항▼
1. A kit for carrying out a nucleic acid amplification reaction, wherein said kit comprises a pair of primers, wherein at least one primer of said pair is reversibly blocked by the covalent attachment of a triarylmethyl group to the 3' terminal OH. 2. A kit of claim 1, wherein said triarylmethyl gro
1. A kit for carrying out a nucleic acid amplification reaction, wherein said kit comprises a pair of primers, wherein at least one primer of said pair is reversibly blocked by the covalent attachment of a triarylmethyl group to the 3' terminal OH. 2. A kit of claim 1, wherein said triarylmethyl group has the formula: wherein R1,R2,and R3represent independently an aryl group; a nitrogen, sulfur, or oxygen-containing heterocyclic ring; or an aryl group with a monosubstituent selected from the group consisting of halide, nitro, lower alkyl, lower alkoxy, and aryl, aralkyl, and cycloalkyl containing up to 10 carbon atoms; wherein R2and R3each also may be alkyl, aralkyl, or cycloalkyl containing up to 10 carbon atoms. 3. A kit of claim 1, wherein said triarylmethyl group is dimethoxytrityl. 4. A kit of claim 3, wherein both primers of said pair of primers are reversibly blocked. 5. A method for amplifying a nucleic acid target sequence, wherein said method comprises carrying out an amplification reaction using a pair of primers, wherein at least one primer of said pair is reversibly blocked by the covalent attachment of a triarylmethyl group to the 3' terminal OH. 6. A method of claim 5, wherein said triarylmethyl group has the formula: wherein R1,R2,and R3represent independently an aryl group; a nitrogen, sulfur, or oxygen-containing heterocyclic ring; or an aryl group with a monosubstituent selected from the group consisting of halide, nitro, lower alkyl, lower alkoxy, and aryl, aralkyl, and cycloalkyl containing up to 10 carbon atoms; wherein R2and R3each also may be alkyl, aralkyl, or cycloalkyl containing up to 10 carbon atoms. 7. A method of claim 5, wherein said triarylmethyl group is dimethoxytrityl. 8. A method of claim 7, wherein both primers of said pair of primers are reversibly blocked.
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이 특허에 인용된 특허 (4)
Will Stephen Gordon, Modified nucleic acid amplification primers.
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Moser, Michael James; Marshall, David J.; Prudent, James R.; Van Hout, Christopher V.; Larsen, Christine A., Polymerase inhibitor and method of using same.
Moser, Michael James; Marshall, David J.; Prudent, James R.; Van Hout, Cristopher V.; Larsen, Christine A., Polymerase inhibitor and method of using same.
Martin, Patrick K.; Simpson, David A.; Hardy, Christine D., Polypeptides having nucleic acid binding activity and compositions and methods for nucleic acid amplification.
Althaus, John S.; Kyonghoon, Lee; Namasivayam, Vijay; Brahmasandra, Sundaresh N.; Handique, Kalyan, System and method for electrochemical detection of biological compounds.
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