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    • 4. 发明申请
      WO2004094662A3 METHOD OF PREPARING DNA FRAGMENTS BY SELECTIVE FRAGMENTATION OF NUCLEIC ACIDS AND APPLICATIONS THEREOF 审中-公开
    • METHOD OF PREPARING DNA FRAGMENTS BY SELECTIVE FRAGMENTATION OF NUCLEIC ACIDS AND APPLICATIONS THEREOF
    • 通过核酸选择性分解制备DNA片段的方法及其应用
    • WO2004094662A3
    • 2005-04-14
    • PCT/FR2004000963
    • 2004-04-19
    • COMMISSARIAT ENERGIE ATOMIQUE , UNIV JOSEPH FOURIER , CENTRE NAT RECH SCIENT , BRACHET ANNE-GAELLE , RIZO PHILIPPE , TABERLET PIERRE , TEXIER-NOGUES ISABELLE
    • BRACHET ANNE-GAELLE , RIZO PHILIPPE , TABERLET PIERRE , TEXIER-NOGUES ISABELLE
    • C12Q1/68
    • C12Q1/6855 , C12Q2525/131 , C12Q2521/313
    • The invention relates to a method of preparing DNA fragments by selective fragmentation of nucleic acid fragments. The inventive method comprises a first selection step involving selection of short fragments, consisting in: a) preparing first double-stranded DNA fragments F1 using at least one restriction enzyme E1 which can randomly fragment the nucleic acid sample to be analysed, by generating said DNA fragments F1 with blunt or cohesive ends; b) ligating the ends of the DNA fragments F1 obtained in step (a) to at least one adapter AA'; c) cleaving the DNA fragments F'1 obtained in step (b) using a restriction enzyme E2, such as to select a fraction of short fragments F2; and d) using any suitable means to purify the aforementioned fraction of short fragments F2. The inventive method also comprises the following optional step involving the second selection of one or more fragment sub-groups from the fraction of short fragments F2 obtained in step (d), said optional step consisting in: e) ligating the free end of the short fragments F2 obtained in step d) to at least one second complementary adapter BB' (production of fragments F'2); and f) amplifying short fragments F'2. The invention also relates to the applications of the above-mentioned method for the analysis of genomes and transcriptomes.
    • 本发明涉及通过选择性片段化核酸片段制备DNA片段的方法。 本发明的方法包括涉及选择短片段的第一选择步骤，其包括：a）使用至少一种限制酶E1制备第一双链DNA片段F1，所述限制酶E1可以通过产生所述DNA产生随机片段化的待分析的核酸样品 片段F1具有钝或粘性末端; b）将步骤（a）中获得的DNA片段F1的末端连接至至少一个衔接子AA'; c）使用限制酶E2切割步骤（b）中获得的DNA片段F'1，以选择一小部分短片段F2; 和d）使用任何合适的方法来纯化上述部分的短片段F2。 本发明的方法还包括以下任选步骤，其涉及从步骤（d）中获得的短片段F2的一个或多个片段亚组的第二选择，所述任选步骤包括：e）连接短片段的自由末端 步骤d）中获得的片段F2至少一个第二互补适配体BB'（片段F'2的产生）; 和f）放大短片段F'2。 本发明还涉及上述用于分析基因组和转录组的方法的应用。
    • 基本信息 添加关注 加入工作空间 PDF全文 PDF下载 全文下载 相似专利 双屏查看 权利要求书 说明书全文 Global Dossier Espacenet 法律信息 同族专利 专利引用
    • 6. 发明专利
      AT415493T  未知
    • AT415493T
    • 2008-12-15
    • AT04742541
    • 2004-04-19
    • COMMISSARIAT ENERGIE ATOMIQUE , UNIV JOSEPH FOURIER , CENTRE NAT RECH SCIENT
    • BRACHET ANNE-GAELLE , RIZO PHILIPPE , TABERLET PIERRE , TEXIER-NOGUES ISABELLE
    • C12Q1/68
    • Method for preparing fragments (F) of DNA from a sample of nucleic acids being analyzed, by selective fragmentation. Method for preparing fragments (F) of DNA from a sample of nucleic acids being analyzed, by selective fragmentation comprises at least one of the steps: (1) a first selection of short fragments by producing first double-stranded DNA fragments (F1), using at least one restriction enzyme (E1) that cuts in a random manner to generate free or cohesive ends; (2) ligating the ends of F1 to an adapter (AA') to form a motif (M) (at the junction of the complementary strand of AA' and the 5'-end of F1) where the sequence of (M) is N-x first base pairs (bp) from the recognition site (of N bp) of a restriction enzyme (E2) for which the cutting site is downstream of the recognition site (x is between 1 and N+1), and the 3'-end (situated in the 5'-region of F1) is the restriction site for E1, resulting in fragments F'1; (3) cutting F'1 close to their 5'-ends, using E2, to select a fraction of short fragments F2 and these are purified by any standard method; (4) a second selection of one or more sub-sets of F2 by ligating the free end (not bound to AA') to a second complementary adapter (BB') to form fragments F'2; (5) amplifying F'2 linked to both adapters, using appropriate primers, at least one being optionally 5'-labeled, so as to select at least one subset of F'2. Independent claims are also included for the following: (1) DNA fragment (A1) that represents a genetic marker and prepared by the new method; (2) DNA chip that carries (A1); (3) method for hybridization of nucleic acid, using (A1) or the chip of (2); (4) kit for method (3) containing (A1) or the chip of (2); and (5) kit for performing the new method that contains at least one adapter AA'.
    • 基本信息 添加关注 加入工作空间 PDF全文 PDF下载 全文下载 相似专利 双屏查看 权利要求书 说明书全文 Espacenet 法律信息 同族专利 专利引用
    • 8. 发明专利
      DE602004017980D1  未知
    • DE602004017980D1
    • 2009-01-08
    • DE602004017980
    • 2004-04-19
    • COMMISSARIAT ENERGIE ATOMIQUE , UNIV JOSEPH FOURIER , CENTRE NAT RECH SCIENT
    • BRACHET ANNE-GAELLE , RIZO PHILIPPE , TABERLET PIERRE , TEXIER-NOGUES ISABELLE
    • C12Q1/68
    • Method for preparing fragments (F) of DNA from a sample of nucleic acids being analyzed, by selective fragmentation. Method for preparing fragments (F) of DNA from a sample of nucleic acids being analyzed, by selective fragmentation comprises at least one of the steps: (1) a first selection of short fragments by producing first double-stranded DNA fragments (F1), using at least one restriction enzyme (E1) that cuts in a random manner to generate free or cohesive ends; (2) ligating the ends of F1 to an adapter (AA') to form a motif (M) (at the junction of the complementary strand of AA' and the 5'-end of F1) where the sequence of (M) is N-x first base pairs (bp) from the recognition site (of N bp) of a restriction enzyme (E2) for which the cutting site is downstream of the recognition site (x is between 1 and N+1), and the 3'-end (situated in the 5'-region of F1) is the restriction site for E1, resulting in fragments F'1; (3) cutting F'1 close to their 5'-ends, using E2, to select a fraction of short fragments F2 and these are purified by any standard method; (4) a second selection of one or more sub-sets of F2 by ligating the free end (not bound to AA') to a second complementary adapter (BB') to form fragments F'2; (5) amplifying F'2 linked to both adapters, using appropriate primers, at least one being optionally 5'-labeled, so as to select at least one subset of F'2. Independent claims are also included for the following: (1) DNA fragment (A1) that represents a genetic marker and prepared by the new method; (2) DNA chip that carries (A1); (3) method for hybridization of nucleic acid, using (A1) or the chip of (2); (4) kit for method (3) containing (A1) or the chip of (2); and (5) kit for performing the new method that contains at least one adapter AA'.
    • 基本信息 添加关注 加入工作空间 PDF全文 PDF下载 全文下载 相似专利 双屏查看 权利要求书 说明书全文 Espacenet 法律信息 同族专利 专利引用
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