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    Method for making full-length cDNA libraries
    1.
    发明授权
    Method for making full-length cDNA libraries 失效
    制备全长cDNA文库的方法

    公开(公告)号：US06174669B1

    公开(公告)日：2001-01-16

    申请号：US08752540

    申请日：1996-11-20

    申请人： Yoshihide Hayashizaki ,  Piero Carninci ,  Claudio Schneider

    发明人： Yoshihide Hayashizaki ,  Piero Carninci ,  Claudio Schneider

    IPC分类号： C12Q168

    CPC分类号： C07H21/00 ,  C07B2200/11 ,  C12N15/1093 ,  C12N15/1096 ,  C40B40/00

    摘要： Disclosed is a method for making full-length CDNA libraries, which is for making libraries of cDNAs having a length corresponding to a full-length of mRNAs and comprises the following steps of; binding a tag molecule to a diol structure present in 5′ Cap (7MeGpppN) sites of mRNAs, forming RNA-DNA hybrids by reverse transcription using primers such as oligo dT and the mRNAs connected with the tag molecule as templates, and separating RNA-DNA hybrids carrying a DNA corresponding to a full-length of mRNAs from the RNA-DNA hybrids formed above by using function of the tag molecule. To obtain mRNA connected with a tag molecule, the diol structure present in 5′ Cap site of mRNA is subjected to a ring-open reaction by oxidation with sodium periodate to form a dialdehyde and the dialdehyde is reacted with a tag molecule having a hydrazine terminus. According to the present invention, there are provided a novel method capable of efficiently labeling 5′ Cap site and a method for making full-length cDNA libraries utilizing the labeling method.

    摘要翻译： 公开了一种制备全长CDNA文库的方法，其用于制备具有对应于全长mRNA的长度的cDNA文库，并且包括以下步骤： 将标签分子与存在于mRNA的5'帽（7MeGpppN）位点的二醇结构结合，通过使用诸如oligo dT的引物和与标签分子连接的mRNA作为模板通过逆转录形成RNA-DNA杂交体，并分离RNA-DNA 通过使用标签分子的功能，携带与来自上述RNA-DNA杂交体的全长mRNA相对应的DNA的杂交体。 为了获得与标签分子连接的mRNA，存在于mRNA的5'帽位点的二醇结构通过用高碘酸钠氧化进行开环反应以形成二醛，并且使二醛与具有肼末端的标签分子反应 。 根据本发明，提供了一种能够利用标记方法有效地标记5'帽位点和制备全长cDNA文库的方法的新方法。

    Extraction of genomic DNA from blood using cationic detergents
    2.
    发明授权
    Extraction of genomic DNA from blood using cationic detergents 失效
    使用阳离子洗涤剂从血液中提取基因组DNA

    公开(公告)号：US5596092A

    公开(公告)日：1997-01-21

    申请号：US268577

    申请日：1994-07-06

    申请人： Claudio Schneider

    发明人： Claudio Schneider

    IPC分类号： C12N15/09 ,  C12N15/10 ,  C07H21/00

    CPC分类号： C12N15/1003 ,  C12N15/1017

    摘要： A process is disclosed for extracting genomic DNA from a biological sample containing a cell source comprising lysing the biological sample with a first cationic detergent in a solution of high salt concentration greater than 0.5M, under heating at 40.degree. to 80.degree. C., so as to solubilize nucleic acids and proteins present in the cell source. The lysed solution is then extracted with an organic solvent by leaving the DNA in the aqueous phase and precipitating any protein present in the aqueous phase. The DNA is then precipitated from the aqueous solution by diluting it at a salt concentration of below 0.5M and adding thereto a second cationic detergent.

    摘要翻译： 公开了一种用于从含有细胞源的生物样品中提取基因组DNA的方法，所述生物样品包括在40℃至80℃加热下在高盐浓度大于0.5M的溶液中用第一阳离子洗涤剂裂解生物样品，因此 以溶解细胞源中存在的核酸和蛋白质。 然后将溶解的溶液用有机溶剂萃取，将DNA留在水相中并沉淀存在于水相中的任何蛋白质。 然后通过在低于0.5M的盐浓度下稀释其从水溶液中沉淀DNA并向其中加入第二阳离子洗涤剂。