Support bound probes and methods of analysis using the same
원문보기
IPC분류정보
국가/구분
United States(US) Patent
등록
국제특허분류(IPC7판)
C12Q-001/68
C07H-021/02
출원번호
US-0557875
(2000-04-24)
발명자
/ 주소
Fodor, Stephen P. A.
Pirrung, Michael C.
Stryer, Lubert
Read, J. Leighton
출원인 / 주소
Affymetrix, Inc.
대리인 / 주소
Townsend and Townsend and Crew LLP
인용정보
피인용 횟수 :
73인용 특허 :
203
초록▼
The present invention provides methods and apparatus for sequencing, fingerprinting and mapping biological macromolecules, typically biological polymers. The methods make use of a plurality of sequence specific recognition reagents which can also be used for classification of biological samples, and
The present invention provides methods and apparatus for sequencing, fingerprinting and mapping biological macromolecules, typically biological polymers. The methods make use of a plurality of sequence specific recognition reagents which can also be used for classification of biological samples, and to characterize their sources.
대표청구항▼
The present invention provides methods and apparatus for sequencing, fingerprinting and mapping biological macromolecules, typically biological polymers. The methods make use of a plurality of sequence specific recognition reagents which can also be used for classification of biological samples, and
The present invention provides methods and apparatus for sequencing, fingerprinting and mapping biological macromolecules, typically biological polymers. The methods make use of a plurality of sequence specific recognition reagents which can also be used for classification of biological samples, and to characterize their sources. ng cardiac hypertrophy from a tissue sample, comprising hybridizing the probe of claim 23 with nucleic acid in the sample, said hybridization is indicative of the presence of SEQ ID NO:5. 28. A method of detecting cardiac hypertrophy from a tissue sample, comprising hybridizing the probe of claim 24 with nucleic acid in the sample, said hybridization is indicative of the presence of SEQ ID NO:8. idize to opposite strands of a target DNA or RNA and which each have a transcriptase or polymerase binding site. 6. A kit for use in RT-PCR amplification techniques, comprising: a) a reversibly inactivated thermostable reverse transcriptase, wherein the transcriptase is in the form of a fusion protein that is attached to an immobilizing moiety via a pair of affinity binding partners, wherein binding of the fusion protein to the immobilizing moiety reversibly inactivates the reverse transcriptase, and release of the fusion protein from the immobilizing moiety activates the transcriptase, and wherein one partner of the pair of affinity binding partners is attached to or is inherently part of the immobilizing moiety, and the other partner of the pair of affinity binding partners is attached to or is inherently part of the fusion protein; b) a RT primer which binds to a target RNA for generating a complementary DNA (cDNA); and c) a pair of PCR primers which hybridize to opposite strands of a double-stranded DNA, one strand of which is the cDNA. 7. A kit for use in Q-beta amplification techniques, comprising: a) reversibly inactivated a thermostable RNA-directed RNA polymerase, wherein the polymerase is in the form of a fusion protein that is bound to an immobilizing moiety via a pair of affinity binding partners, wherein binding of the fusion protein to the immobilizing moiety reversibly inactivates the polymerase, and release of the fusion protein from the immobilizing moiety activates the polymerase, and wherein one partner of the pair of affinity binding partners is attached to or is inherently part of the immobilizing moiety, and the other partner of the pair of affinity binding partners is attached to or is inherently part of the fusion protein; and b) an RNA probe with a 5'-MDV-1 structure, the RNA probe being immobilized or permitting immobilization. 8. A method of activating a reversibly inactivated enzyme or a fragment thereof having catalytic activity of the enzyme, wherein the enzyme or fragment is in the form of a fusion protein and is inactivated by attachment of the fusion protein to an immobilizing moiety via a pair of affinity binding partners and is activated by release of the fusion protein from the immobilizing moiety, said method comprising releasing the fusion protein from said immobilizing moiety, wherein the enzyme is selected from the group consisting of DNA polymerases, DNA ligases, reverse transcriptases, and RNA polymerases, and wherein one partner of the pair of affinity binding partners is attached to or is inherently part of the immobilizing moiety, and the other partner of the pair of affinity binding partners is attached to or is inherently part of the fusion protein. 9. A method as claimed in claim 8 wherein said enzyme or fragment is thermostable. 10. A method as claimed in claim 9 wherein said enzyme is DNA polymerase from Thermus thermophilus or Thermus aquaticus. 11. A method as claimed in claim 8 wherein said fusion protein is bound to the immobilizing moiety via a heat-labile linkage. 12. A method as claimed in claim 8, wherein one partner of said pair of affinity binding partners is inherently part of said fusion protein. 13. A method as claimed in claim 8, wherein said pair of affinity binding partners is (a) protein A or a part thereof having the function of protein A, and (b) immunoglobulin G (IgG) or a part thereof having the function of IgG. 14. A method as claimed in claim 8, wherein said pair of affinity binding partners is (a) protein G or a part thereof having the function of protein G, and (b) human serum albumin (HSA) or a part thereof having the function of HSA. 15. A method as claimed in claim 12 wherein said one partner is an epitope of said enzyme or fragment, or of a moiety appended to said enzyme or fragment, and the other binding partner is an antibody which specifically recognizes said epitope. 16. A method as claimed in claim 8 wherein said immobilizing moi
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이 특허에 인용된 특허 (203)
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