公开(公告)号：US07238530B2

公开(公告)日：2007-07-03

申请号：US09760085

申请日：2001-01-12

申请人： Jaap Goudsmit ,  Cornelis Johannes Andreas Sol ,  Marcellinus Gualbertus Hubertus Maria Beld ,  Willem René Boom

发明人： Jaap Goudsmit ,  Cornelis Johannes Andreas Sol ,  Marcellinus Gualbertus Hubertus Maria Beld ,  Willem René Boom

IPC分类号： C12Q1/68 ,  C07H21/02 ,  C07H19/04

CPC分类号： C12N15/1006 ,  C12N15/10 ,  C12Q1/6806 ,  C12Q1/6855 ,  C12Q2527/137 ,  C12Q2527/125

摘要： The invention relates to methods for easily separating single stranded nucleic acid material from double stranded nucleic acid material in a sample containing both. By the right choice of at lease one chaotropic agent, preferably a guanidine salt, at a selected concentration and other suitable conditions such a chelating agents, pH and the like, it is possible to bind double stranded material to a solid phase such as silica particles, whereas single stranded material will not bind under those circumstances. By separating the silica particles from the sample the double stranded nucleic acid material is removed. It can easily be eluted from the silica particles. In a second step the single stranded material may be bound to a solid phase by selecting a different set of conditions. The particles can again be separated from the sample and the single stranded material may now be eluted. For very efficent separations, the process may be repeated. Following the separation of the two kinds of nucleic acid, either kind may be amplified. Methods of amplification are provided which do not need sequence data of the material to be amplified. In these methods a primer will be provided with an amplification motif and a random hybridization motif.

摘要翻译： 本发明涉及在含有两者的样品中容易地从双链核酸材料中分离单链核酸材料的方法。 通过正确选择一种离液剂，优选胍盐，以选定的浓度和其它合适的条件如螯合剂，pH等，可以将双链材料结合到固相如二氧化硅颗粒 而单链材料在这种情况下不会结合。 通过从样品中分离二氧化硅颗粒，除去双链核酸材料。 它可以容易地从二氧化硅颗粒中洗脱出来。 在第二步中，单链材料可以通过选择不同的条件集合而与固相结合。 颗粒可以再次与样品分离，单链材料现在可以被洗脱。 为了非常有效的分离，可以重复该过程。 在分离两种核酸后，可以扩增任一种。 提供了不需要放大材料的序列数据的扩增方法。 在这些方法中，引物将具有扩增基序和随机杂交基序。