公开(公告)号：US5346999A

公开(公告)日：1994-09-13

申请号：US328471

申请日：1989-03-24

申请人： Guy R. Cathcart ,  Paul D. Grossman ,  P. Eric Mayrand ,  Eric S. Nordman ,  Norman M. Whiteley

发明人： Guy R. Cathcart ,  Paul D. Grossman ,  P. Eric Mayrand ,  Eric S. Nordman ,  Norman M. Whiteley

IPC分类号： B01J19/00 ,  C12M1/00 ,  C12N9/06 ,  C12N15/10 ,  C40B60/14 ,  C07H23/00

CPC分类号： C12N9/0012 ,  B01J19/0046 ,  C12M47/06 ,  C12M47/12 ,  C12N15/1003 ,  B01J2219/00286 ,  B01J2219/00308 ,  B01J2219/00331 ,  B01J2219/00389 ,  B01J2219/00488 ,  B01J2219/00689 ,  B01J2219/00695 ,  C40B60/14 ,  Y10T436/25125 ,  Y10T436/25375

摘要： An automated apparatus is provided which implements a new method of extracting and purifying nucleic acids from cells without the use of centrifugation. In the method, a lysate is created by treating the cells with proteinase K in the presence of a lysis buffer having a high concentration of a salt. The lysate is mixed with a phenol-based solvent system, thereby creating an emulsion. The emulsion is heated to promote phase separation. Similarly, the rate of phase separation is also enhanced by increasing the surface area of the emulsion. Once the phase separation is complete, the lower organic phase is removed and the upper aqueous phase is repeatedly extracted with the phenol-based solvent a preselected number of times, and is finally extracted using chloroform. The remaining aqueous phase is then dialyzed to further purify and concentrate the nucleic acid solution. Two preferred embodiments of apparatus are presented to accomplish this extraction.

摘要翻译： 提供了一种自动化装置，其实现了从细胞中提取和纯化核酸而不使用离心的新方法。 在该方法中，通过在具有高浓度盐的裂解缓冲液存在下用蛋白酶K处理细胞产生裂解物。 将裂解物与苯酚基溶剂体系混合，从而产生乳液。 将乳液加热以促进相分离。 类似地，通过增加乳液的表面积也增加了相分离速率。 一旦相分离完成，除去较低的有机相，并用苯酚类溶剂预选次数反复提取上部水相，最后用氯仿萃取。 然后将剩余的水相透析以进一步净化并浓缩核酸溶液。 呈现装置的两个优选实施例以完成该提取。