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    • 5. 发明授权
    • Use of class IIB restriction endonucleases in 2nd generation sequencing applications
    • 在第二代测序应用中使用IIB类限制性内切核酸酶
    • US08980551B2
    • 2015-03-17
    • US13318278
    • 2010-05-05
    • Norman WarthmannDetlef Weigel
    • Norman WarthmannDetlef Weigel
    • C12Q1/68C12P19/34C07H21/00C07H21/02C07H21/04C12N15/64C12N15/10C12N15/66
    • C12N15/64C12N15/1065C12N15/66
    • The present invention relates to a method for genotyping DNA molecules contained in at least one DNA sample. The method includes: (a) digesting the DNA molecules contained in at least one DNA sample with a class IIB restriction endonuclease to generate DNA fragments; (b) optionally separating DNA fragments comprising the recognition site for the class IIB restriction endonuclease from the remaining DNA fragments; (c) attaching at least one adaptor DNA to the 5′ and/or 3′ end of one or both strands of the DNA fragments comprising the recognition site for the class IIB restriction endonuclease obtained in a) or separated in b) to form adaptor-fragment constructs; (d) determining the sequence of at least a fraction of the DNA fragments obtained in c); and (e) assigning genotypes to the at least one DNA sample analyzed based on the sequence data obtained in d). The present invention further relates to method for determining the position of DNA molecules comprised in a DNA library within the DNA sequence represented by the DNA library or within a known DNA sequence and for establishing a cross-reference between individual DNA molecules and their location in an at least three dimensional matrix.
    • 本发明涉及一种用于对包含在至少一个DNA样品中的DNA分子进行基因分型的方法。 该方法包括:(a)用IIB限制性内切核酸酶消化至少一个DNA样品中所含的DNA分子以产生DNA片段; (b)任选地从其余DNA片段分离包含IIB限制性内切核酸酶的识别位点的DNA片段; (c)将至少一个接头DNA连接到包含a)中获得的IIB限制性内切核酸酶的识别位点或在b)中分离的DNA片段的一条或两条链的5'和/或3'末端,以形成衔接子 片段构造; (d)确定在c)中获得的DNA片段的至少一部分的序列; 和(e)基于在d)中获得的序列数据将基因型分配给所分析的至少一个DNA样品。 本发明还涉及确定由DNA文库或已知DNA序列表示的DNA序列内的DNA文库中包含的DNA分子的位置的方法,并且用于在单个DNA分子与其位置之间建立交叉引用 至少三维矩阵。
    • 7. 发明申请
    • THE USE OF CLASS IIB RESTRICTION ENDONUCLEASES IN 2ND GENERATION SEQUENCING APPLICATIONS
    • IIB限制性内切酶在第二代序列应用中的应用
    • US20120094847A1
    • 2012-04-19
    • US13318278
    • 2010-05-05
    • Norman WarthmannDetlef Weigel
    • Norman WarthmannDetlef Weigel
    • C40B20/00C12P19/34C12Q1/68
    • C12N15/64C12N15/1065C12N15/66
    • The present invention relates to a method for genotyping DNA molecules contained in at least one DNA sample comprising: (a) digesting the DNA molecules contained in at least one DNA sample with a class IIB restriction endonuclease to generate DNA fragments; (b) optionally separating DNA fragments comprising the recognition site for said class IIB restriction endonuclease from the remaining DNA fragments; (c) attaching at least one adaptor DNA to the 5′ and/or 3′ end of one or both strands of the DNA fragments comprising the recognition site for said class IIB restriction endonuclease obtained in a) or separated in b) to form adaptor-fragment constructs; (d) determining the sequence of at least a fraction of the DNA fragments obtained in c); and (e) assigning genotypes to said at least one DNA sample analysed based on the sequence data obtained in d). The present invention further relates to method for determining the position of DNA molecules comprised in a DNA library within the DNA sequence represented by said DNA library or within a known DNA sequence and for establishing a cross-reference between individual DNA molecules and their location in an at least three dimensional matrix.
    • 本发明涉及一种用于对包含在至少一个DNA样品中的DNA分子进行基因分型的方法,包括:(a)用IIB限制性内切核酸酶消化至少一个DNA样品中所含的DNA分子以产生DNA片段; (b)任选地将包含所述IIB限制性内切核酸酶的识别位点的DNA片段与剩余的DNA片段分离; (c)将至少一个接头DNA连接到包含a)所获得的所述IIB限制性内切核酸酶的识别位点或在b)中分离的DNA片段的一条或两条链的5'和/或3'末端,以形成衔接子 片段构造; (d)确定在c)中获得的DNA片段的至少一部分的序列; 和(e)基于在d)中获得的序列数据将基因型分配给所分析的所述至少一个DNA样品。 本发明还涉及用于确定由所述DNA文库或已知DNA序列表示的DNA序列内的DNA文库中包含的DNA分子的位置的方法,并且用于在单个DNA分子与其位置之间建立交叉参照 至少三维矩阵。