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    METHODS FOR AMPLIFYING POLYMERIC NUCLEIC ACIDS
    4.
    发明公开
    METHODS FOR AMPLIFYING POLYMERIC NUCLEIC ACIDS 有权
    方法扩增高分子NUCLEIC

    公开(公告)号：EP1649065A2

    公开(公告)日：2006-04-26

    申请号：EP04779936.6

    申请日：2004-08-02

    申请人： INTEGRATED DNA TECHNOLOGIES, INC.

    发明人： BEHLKE, Mark, Aaron ,  WALDER, Joseph, Alan ,  MANTHEY, Jeffrey, A.

    IPC分类号： C12Q1/68 ,  C12P19/34 ,  C07H21/04

    CPC分类号： C12Q1/6848 ,  C12Q1/6853 ,  C12Q2549/119 ,  C12Q2531/101 ,  C12Q2525/186 ,  C12Q2525/121 ,  C12Q2531/113

    摘要： The invention provides compositions and methods for amplifying nucleic acid polymer sequences in a high complexity nucleic acid sample. The unique compositions of the invention include a primer set composed of a mixture of two types of primers for DNA synthesis. For extension in one direction, the primers all contain modifications that destroy their ability to serve as templates that can be copied by DNA polymerases. For extension in the opposite direction the set includes at least one primer that can serve as a template and be replicated by DNA polymerases throughout its length. The method can be carried out by mixing the nucleic acid polymer sequence of interest with the set of DNA synthesis primers in an amplification reaction mixture. The reaction mixture is then subjected to temperature cycling analogous to the temperature cycling in PCR reactions. At least one primer in the primer set hybridizes to the nucleic acid polymer. It is preferred that the non-replicable primer hybridizes to the nucleic acid polymer and is extended to produce an extension product that contains sequence from the nucleic acid polymer to which the replicable primer then hybridizes. Of course, if the nucleic acid polymer is double stranded, both the replicable and nonreplicable primers will hybridize and be extended by DNA polymerase.

    METHODS AND SYSTEMS FOR ESTIMATING THE MELTING TEMPERATURE ( i T /i sb i M /i /sb ) FOR POLYNUCLEOTIDE MOLECULES
    5.
    发明公开
    METHODS AND SYSTEMS FOR ESTIMATING THE MELTING TEMPERATURE ( i T /i sb i M /i /sb ) FOR POLYNUCLEOTIDE MOLECULES 有权
    METHOD AND SYSTEM FOR FOR多核苷酸估算的熔融温度（T M）

    公开(公告)号：EP1543438A2

    公开(公告)日：2005-06-22

    申请号：EP03754518.3

    申请日：2003-09-12

    申请人： INTEGRATED DNA TECHNOLOGIES, INC.

    发明人： OWCZARZY, Richard ,  WALDER, Joseph, Alan ,  HUANG, Lingyan ,  BEHLKE, Mark, Aaron

    IPC分类号： G06F17/00 ,  G06F17/10 ,  G06F17/11 ,  C12Q1/68

    CPC分类号： G06F19/20 ,  C12Q1/6813 ,  G06F19/22

    摘要： The invention relates to methods and systems for predicting or estimating the melting temperature of duplex nucleic acids, particularly duplexes of oligonucleotides which may be used, for example, as primers or probes in PCR and/or hybridization assays. The invention also relates to methods and systems for designing and selecting oligonucleotide probes and primers having a predicted melting temperature which is optimized for such assays. To this end, algorithms and methods are provided for predicting the melting temperature of a nucleic acid having a predetermined sequence. These methods and algorithms estimate the melting temperature of a nucleic acid duplex under particular salt conditions. The methods and algorithms use novel formulas, having terms and coefficients that are functions of the particular nucleotide sequence, to estimate the effect of particular salt conditions on the melting temperature. As such, the methods and systems of the invention provide superior result compared to existing methods, which do not consider sequence dependent effects of changing salt conditions. ­

    RNASE H-BASED ASSAYS UTILIZING MODIFIED RNA MONOMERS
    8.
    发明公开
    RNASE H-BASED ASSAYS UTILIZING MODIFIED RNA MONOMERS 审中-公开
    测试AUF RNASE-H-BASIS UNTER VERWENDUNG MODIFIZIERTER RNA-MONOMERE

    公开(公告)号：EP2971072A1

    公开(公告)日：2016-01-20

    申请号：EP13811693.4

    申请日：2013-12-02

    申请人： Integrated DNA Technologies Inc.

    发明人： WALDER, Joseph, Alan ,  BEHLKE, Mark, Aaron ,  ROSE, Scott, D. ,  DOBOSY, Joseph, R. ,  RUPP, Susan, M.

    IPC分类号： C12Q1/68

    CPC分类号： C12Q1/6853

    摘要： Methods and compositions are provided for improving specificity during amplification of a target DNA sequence. The methods and compositions rely upon the use of an RNase H enzyme, a polymerase, and RNase H enzyme-sensitive, blocked-cleavable oligonucleotide primers in the amplification reactions, wherein the reaction mixtures include either an optimized final concentration of a divalent metal salt comprising 2.0 mM or less of free Mg
    ++ cation and/or an optimized final concentration of a non-ionic detergent comprising at least about 0.001% polyethylene glycol hexadecyl ether.

    摘要翻译： 提供了用于在靶DNA序列扩增期间提高特异性的方法和组合物。 所述方法和组合物依赖于在扩增反应中使用RNase H酶，聚合酶和RNase H酶敏感的可封闭切割的寡核苷酸引物，其中反应混合物包括优化的最终浓度的二价金属盐，其包含 2.0mM或更少的游离Mg ++阳离子和/或包含至少约0.001％聚乙二醇十六烷基醚的非离子洗涤剂的优化最终浓度。