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1. 发明申请

WO2004085678A3 METHOD OF PREPARING DNA FRAGMENTS AND APPLICATIONS THEREOF 审中-公开
标题翻译：制备DNA片段的方法及其应用
公开(公告)号：WO2004085678A3
公开(公告)日：2004-12-09
申请号：PCT/FR2004000671
申请日：2004-03-18
申请人： COMMISSARIAT ENERGIE ATOMIQUE , BRACHET ANNE-GAELLE , RIZO PHILIPPE
发明人： BRACHET ANNE-GAELLE , RIZO PHILIPPE
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6855 , C12Q2525/131
摘要： The invention relates to a method of preparing DNA fragments and to the applications thereof, in particular for the hybridisation of nucleic acids. The inventive method is essentially characterised in that it consists of at least the following steps comprising: (a) preparation of double-stranded DNA fragments from a sample of nucleic acids to be analysed; (b) ligation of the ends of the aforementioned DNA fragments to a double-stranded oligonucleotide adapter (adapter AA') comprising the site for the recognition of a restriction enzyme of which the cleavage site is situated downstream of said recognition site; (c) amplification of the fragments linked to the above-mentioned adapter, using a pair of suitable primers, one of which is optionally marked at the 5' end thereof; and (d) cleavage of said DNA fragments close to one of the ends of same, using the restriction enzyme, such as to generate short fragments.
摘要翻译：本发明涉及制备DNA片段的方法及其应用，特别是用于核酸杂交的方法。本发明的方法的基本特征在于其至少由以下步骤组成，所述步骤包括：（a）从待分析的核酸样品制备双链DNA片段; （b）将上述DNA片段的末端连接至包含识别限制性酶的位点的双链寡核苷酸接头（接头AA'），其中切割位点位于所述识别位点的下游; （c）使用一对合适的引物扩增与上述接头连接的片段，其中一个任选在其5'末端标记; 和（d）使用限制性内切酶切割靠近其一端的所述DNA片段，以产生短片段。

2. 发明申请

WO2004094662A3 METHOD OF PREPARING DNA FRAGMENTS BY SELECTIVE FRAGMENTATION OF NUCLEIC ACIDS AND APPLICATIONS THEREOF 审中-公开
标题翻译：通过核酸选择性分解制备DNA片段的方法及其应用
公开(公告)号：WO2004094662A3
公开(公告)日：2005-04-14
申请号：PCT/FR2004000963
申请日：2004-04-19
申请人： COMMISSARIAT ENERGIE ATOMIQUE , UNIV JOSEPH FOURIER , CENTRE NAT RECH SCIENT , BRACHET ANNE-GAELLE , RIZO PHILIPPE , TABERLET PIERRE , TEXIER-NOGUES ISABELLE
发明人： BRACHET ANNE-GAELLE , RIZO PHILIPPE , TABERLET PIERRE , TEXIER-NOGUES ISABELLE
IPC分类号： C12Q1/68
CPC分类号： C12Q1/6855 , C12Q2525/131 , C12Q2521/313
摘要： 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.
摘要翻译：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。

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