公开(公告)号：US20020086326A1

公开(公告)日：2002-07-04

申请号：US10041890

申请日：2002-01-07

Applicant: Promega Corporation

Inventor： Craig E. Smith ,  Charles K. York

IPC: C12Q001/68 ,  C12N015/00

CPC classification number: C12N15/1013 ,  C12Q1/6806 ,  Y10S977/838 ,  Y10S977/904 ,  Y10S977/916 ,  Y10S977/92 ,  Y10S977/924 ,  Y10S977/96

Abstract: The present invention provides methods for isolating biological target materials, particularly nucleic acids, such as DNA or RNA or hybrid molecules of DNA and RNA, from other substances in a medium using silica magnetic particles. The methods of the present invention involve forming a complex of the silica magnetic particles and the biological target material in a mixture of the medium and particles, separating the complex from the mixture using external magnetic force, and eluting the biological target material from the complex. The preferred embodiments of magnetic silica particles used in the methods and kits of the present invention are capable of forming a complex with at least 2 nullg of biological target material per milligram of particle, and of releasing at least 60% of the material from the complex in the elution step of the method. The methods of the present invention produce isolated biological target material which is substantially free of contaminants, such as metals or macromolecular substances, which can interfere with further processing or analysis, if present.

Abstract translation: 本发明提供了使用二氧化硅磁性颗粒从介质中的其他物质分离生物靶材料，特别是DNA或RNA或DNA和RNA的混合分子的方法。 本发明的方法包括在介质和颗粒的混合物中形成二氧化硅磁性颗粒和生物目标材料的复合物，使用外部磁力从混合物中分离复合物，并从复合物中洗脱生物目标物质。 在本发明的方法和试剂盒中使用的磁性二氧化硅颗粒的优选实施方案能够与每毫克颗粒至少2个生物靶材料形成复合物，并且从复合物中释放至少60％的材料 在该方法的洗脱步骤中。 本发明的方法产生基本上不含污染物（例如金属或大分子物质）的分离的生物靶材料，如果存在可能会干扰进一步的加工或分析。

公开(公告)号：US20040086930A1

公开(公告)日：2004-05-06

申请号：US10694475

申请日：2003-10-27

Applicant: Promega Corporation

Inventor： Allan M. Tereba ,  Rex M. Bitner ,  Susan C. Koller ,  Craig E. Smith ,  Daniel D. Kephart ,  Steven J. Ekenberg

IPC: C12Q001/68

CPC classification number: C12N15/1013 ,  C12Q1/6806 ,  Y10S977/838 ,  Y10S977/904 ,  Y10S977/916 ,  Y10S977/92 ,  Y10S977/924 ,  Y10S977/96

Abstract: The present invention provides methods for isolating a defined quantity of DNA target material from other substances in a medium. The method may be carried out using a known quantity of a silica-containing solid support, such as silica magnetic particles, having a definable capacity for reversibly binding DNA target material, and DNA target material in excess of the binding capacity of the particles. The methods of the present invention involve forming a complex of the silica magnetic particles and the DNA target material in a mixture of the medium and particles, and separating the complex from the mixture using external magnetic force. The DNA target material may then be eluted from the complex. The quantity of DNA target material eluted may be determined based on a calibration model. The methods of the present invention permit isolation of DNA target material which is within a known quantity range. The methods of the invention eliminate the step of quantitating purified biological samples prior to further processing, such as amplification, Short Tandem Repeat (STR) analysis, and DNA sequencing. Samples of the DNA target materials may be obtained from liquid or solid media, such as liquid blood or paper.

Abstract translation: 本发明提供了将定义量的DNA靶物质与介质中的其它物质分离的方法。 该方法可以使用已知量的具有可定义的可逆结合DNA靶材料的能力的二氧化硅固体载体，例如二氧化硅磁性颗粒和超过颗粒结合能力的DNA靶材料进行。 本发明的方法包括在介质和颗粒的混合物中形成二氧化硅磁性颗粒和DNA靶材料的复合物，并且使用外部磁力将复合物与混合物分离。 然后DNA靶物质可以从络合物中洗脱出来。 可以基于校准模型确定洗脱的DNA靶物质的量。 本发明的方法允许分离已知量范围内的DNA靶材料。 本发明的方法消除了在进一步加工之前定量纯化的生物样品的步骤，例如扩增，短串联重复（STR）分析和DNA测序。 可以从液体或固体培养基（例如液体血液或纸）获得DNA靶材料的样品。

公开(公告)号：US20020001812A1

公开(公告)日：2002-01-03

申请号：US09912045

申请日：2001-07-24

Applicant: Promega Corporation.

Inventor： Craig E. Smith ,  Diana L. Holmes ,  Daniel J. Simpson ,  Jehoshua Katzhendler ,  Rex M. Bitner ,  Josephine C. Grosch

IPC: C12Q001/68 ,  G01N031/00 ,  C07H021/02 ,  A01N057/00 ,  C07H021/04 ,  C12P019/34 ,  A61K031/665

CPC classification number: C12N15/1006 ,  B01J41/20 ,  B01J47/04 ,  C12N15/101 ,  C12N15/1013 ,  Y10S435/803 ,  Y10T436/107497

Abstract: Mixed-bed solid phases are provided, with methods for using such solid phases to isolate target nucleic acids, such as plasmid DNA, chromosomal DNA, RNA, or nucleic acids generated by enzymatic amplification from contaminants, including proteins, lipids, cellular debris, or other nucleic acids. The mixed-bed solid phases of this invention are mixtures of at least two different solid phases, each of which has a capacity to bind to the target nucleic acid under different solution conditions, and the capacity to release the nucleic acid under similar elution conditions. By exchanging solution conditions according to the methods of this invention, one can remove contaminants from the target nucleic acid bound to the mixed-bed solid phase, then elute the target nucleic acid in an elution buffer.