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다음과 같은 기능을 한번의 로그인으로 사용 할 수 있습니다.
NTIS 바로가기다음과 같은 기능을 한번의 로그인으로 사용 할 수 있습니다.
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Kafe 바로가기국가/구분 | United States(US) Patent 등록 |
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국제특허분류(IPC7판) |
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출원번호 | US-0268254 (2014-05-02) |
등록번호 | US-10208341 (2019-02-19) |
발명자 / 주소 |
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출원인 / 주소 |
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대리인 / 주소 |
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인용정보 | 피인용 횟수 : 0 인용 특허 : 351 |
The present invention relates to a novel method for analyzing nucleic acid sequences based on real-time detection of DNA polymerase-catalyzed incorporation of each of the four nucleotide bases, supplied individually and serially in a microfluidic system, to a reaction cell containing a template syst
The present invention relates to a novel method for analyzing nucleic acid sequences based on real-time detection of DNA polymerase-catalyzed incorporation of each of the four nucleotide bases, supplied individually and serially in a microfluidic system, to a reaction cell containing a template system comprising a DNA fragment of unknown sequence and an oligonucleotide primer. Incorporation of a nucleotide base into the template system can be detected by any of a variety of methods including but not limited to fluorescence and chemiluminescence detection. Alternatively, microcalorimetic detection of the heat generated by the incorporation of a nucleotide into the extending template system using thermopile, thermistor and refractive index measurements can be used to detect extension reactions.
1. A method for phase error compensation, comprising: providing a plurality of template nucleic acid strands;adding, in an order, individual types of deoxynucleoside triphosphates serially to the plurality of template nucleic acid strands to perform a series of nucleotide incorporation reactions;aft
1. A method for phase error compensation, comprising: providing a plurality of template nucleic acid strands;adding, in an order, individual types of deoxynucleoside triphosphates serially to the plurality of template nucleic acid strands to perform a series of nucleotide incorporation reactions;after each nucleotide incorporation reaction of the series, detecting a signal transmitted from incorporation of nucleotides resulting from the adding of the deoxynucleoside triphosphates, the signal having an amplitude corresponding to a number of nucleotide incorporations; andaltering the order of the addition of the individual types of deoxynucleoside triphosphates to perform one or more subsequent nucleotide incorporation reactions, thereby compensating for phasing errors due to one or more out-of-phase template nucleic acid strand populations. 2. The method of claim 1, wherein the plurality of template nucleic acid strands are hybridized to a primer. 3. The method of claim 1, wherein the one or more out-of-phase template nucleic acid strand populations comprises a trailing fraction of the template nucleic acid strands that are out of phase relative to an in-phase fraction of the template nucleic acid strands because of extension failures. 4. The method of claim 3, further comprising monitoring the trailing fraction of the template nucleic acid strands to determine where and how to alter the order of addition of the individual types of deoxynucleoside triphosphates so as selectively to extend the trailing fraction of the template nucleic acid strands to bring them back into phase with the in-phase fraction of the template nucleic acid strands. 5. The method of claim 3, wherein altering the order of addition of the individual types of deoxynucleoside triphosphates comprises reversing the order of addition of the individual types of deoxynucleoside triphosphates at arbitrary intervals to bring back into phase about one-third of the trailing fraction of the template nucleic acid strands. 6. The method of claim 1, wherein the one or more out-of-phase template nucleic acid strand populations comprise a leading fraction of the template nucleic acid strands that are out of phase relative to an in-phase fraction of the template nucleic acid strands because of incorrect extensions. 7. The method of claim 6, wherein altering the order of addition of the individual types of deoxynucleoside triphosphates comprises reversing the order of addition of the individual types of deoxynucleoside triphosphates at arbitrary intervals to bring back into phase about two-thirds of the leading fraction of the template nucleic acid strands. 8. The method of claim 3, wherein the one or more out-of-phase template nucleic acid strand populations further comprise a leading fraction of the template nucleic acid strands that are out of phase relative to an in-phase fraction of the template nucleic acid strands because of incorrect extensions. 9. The method of claim 8, wherein altering the order of addition of the individual types of deoxynucleoside triphosphates comprises reversing the order of addition of the deoxynucleoside triphosphates at arbitrary intervals to bring back into phase about two-thirds of the leading fraction of the template nucleic acid strands. 10. A method for phase error compensation, comprising: providing a plurality of template nucleic acid strands;adding, in an order, individual types of deoxynucleoside triphosphates serially to the plurality of template nucleic acid strands to perform a series of nucleotide incorporation reactions;after each nucleotide incorporation reaction of the series, detecting a signal transmitted from incorporation of nucleotides resulting from the adding of the deoxynucleoside triphosphates, the signal having an amplitude corresponding to a number of nucleotide incorporations; andaltering the order of the addition of the individual types of deoxynucleoside triphosphates to perform one or more subsequent nucleotide incorporation reactions, thereby compensating for phasing errors due to one or more out-of-phase template nucleic acid strand populations,wherein the one or more out-of-phase template nucleic acid strand populations comprises a trailing fraction of the template nucleic acid strands that are out of phase relative to an in-phase fraction of the template nucleic acid strands because of extension failures, andwherein altering the order of the addition of the individual types of deoxynucleoside triphosphates is based on monitoring the trailing fraction of the template nucleic acid strands to determine where to alter the order of addition of the deoxynucleoside triphosphates so as selectively to extend the trailing fraction of the template nucleic acid strands to bring them back into phase with the in-phase fraction of the template nucleic acid strands. 11. The method of claim 10, wherein the plurality of template nucleic acid strands are hybridized to a primer. 12. The method of claim 10, wherein altering the order of the addition of the individual types of deoxynucleoside triphosphates comprises reversing the order of addition of the individual types of deoxynucleoside triphosphates at arbitrary intervals to bring back into phase about one-third of the trailing fraction of template nucleic acid strands. 13. A method for phase error compensation, comprising: providing a plurality of template nucleic acid strands;adding, in an order, individual types of deoxynucleoside triphosphates serially to the plurality of template nucleic acid strands to perform a series of nucleotide incorporation reactions;after each nucleotide incorporation reaction of the series, detecting a signal transmitted from incorporation of nucleotides resulting from the adding of the deoxynucleoside triphosphates, the signal having an amplitude corresponding to a number of nucleotide incorporations; andaltering the order of the addition of the individual types of deoxynucleoside triphosphates to perform one or more subsequent nucleotide incorporation reactions, thereby compensating for phasing errors due to one or more out-of-phase template nucleic acid strand populations,wherein the one or more out-of-phase template nucleic acid strand populations comprise a leading fraction of the template nucleic acid strands that are out of phase relative to an in-phase fraction of the template nucleic acid strands because of incorrect extensions; andwherein altering the order of addition of the individual types of deoxynucleoside triphosphates comprises reversing the order of addition of the individual types of deoxynucleoside triphosphates at arbitrary intervals. 14. The method of claim 13, wherein the plurality of template nucleic acid strands are hybridized to a primer.
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