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公开(公告)号：US06521411B2
公开(公告)日：2003-02-18
申请号：US09808447
申请日：2001-03-13
申请人： Karl H. Hecker , Arezou Azarani , David Hornby , Christopher P. Hanna , Douglas T. Gjerde , Maryam Matin
发明人： Karl H. Hecker , Arezou Azarani , David Hornby , Christopher P. Hanna , Douglas T. Gjerde , Maryam Matin
IPC分类号： C12Q168
CPC分类号： C12N15/1096
摘要： In one aspect, the present invention concerns an improved method for the preparation of cDNA libraries. Preferred embodiments of the invention include methods and systems capable of generating cDNA libraries enriched for high molecular weight cDNA inserts. The method generally entails the following steps: (1) size-based separation of a plurality of mRNA molecules by Ion-Pairing Reversed-Phase Chromatography (IP-RPC), preferably using HPLC under denaturing conditions; and (2) collection of a fraction of the mRNA molecules that is enriched for mRNA molecules of a desired size range (in a preferred embodiment of the invention, larger-sized mRNA molecules are collected, e.g., mRNA of length greater than 10 kb). The collected fraction of mRNA molecules can be reverse transcribed to form a library of cDNA inserts enriched for inserts of a desired relative size range.
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公开(公告)号：US06652745B2
公开(公告)日：2003-11-25
申请号：US10047376
申请日：2002-01-14
IPC分类号： B01D1508
CPC分类号： B01D15/325 , B01D15/366 , B01D15/424 , C12N15/101
摘要： An improved separation column and method for separating a mixture of double stranded DNA fragments by Matched Ion Polynucleotide Chromatography. The cylindrical column has an ID greater than about 5 mm and contains polymer beads. The beads have an average diameter of 1 to 100 microns and are unsubstituted polymer beads or are polymer beads substituted with a hydrocarbon moiety having from 1 to 1,000,000 carbons. The preferred beads are characterized by being substantially free from multivalent cations which are free to bind with DNA. The improved column provides enhanced separation of DNA fragments with sizes ranging from about 100 to 20,000 base pairs. The column also provides enhanced separation of heteroduplex and homoduplex DNA molecules in a mutation detection procedure in which the chromatography is performed under conditions effecting partial denaturation of DNA at a site of mismatched base pairs.
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公开(公告)号：US06372142B1
公开(公告)日：2002-04-16
申请号：US09501775
申请日：2000-02-10
IPC分类号： B01D1508
CPC分类号： B01D15/325 , B01D15/366 , B01D15/424 , C12N15/101
摘要： An improved separation column and method for separating a mixture of double stranded DNA fragments by Matched Ion Polynucleotide Chromatography. The cylindrical column has an ID greater than about 5 mm and contains polymer beads. The beads have an average diameter of 1 to 100 microns and are unsubstituted polymer beads or are polymer beads substituted with a hydrocarbon moiety having from 1 to 1,000,000 carbons. The preferred beads are characterized by being substantially free from multivalent cations which are free to bind with DNA. The improved column provides enhanced separation of DNA fragments with sizes ranging from about 100 to 20,000 base pairs. The column also provides enhanced separation of heteroduplex and homoduplex DNA molecules in a mutation detection procedure in which the chromatography is performed under conditions effecting partial denaturation of DNA at a site of mismatched base pairs.
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4.发明申请COMPOSITIONS AND METHODS FOR PREPARING SHORT RNA MOLECULES AND OTHER NUCLEIC ACIDS 审中-公开用于制备短链RNA分子和其他核酸的组合物和方法公开(公告)号：US20090197307A1
公开(公告)日：2009-08-06
申请号：US12189665
申请日：2008-08-11
申请人： Knut R. Madden , Adam N. Harris , Karl H. Hecker , Byung-in Lee
发明人： Knut R. Madden , Adam N. Harris , Karl H. Hecker , Byung-in Lee
CPC分类号： C07H21/02 , B01D15/3804 , C12N15/111 , C12N2330/31
摘要： The invention provides methods of preparing nucleic acids, such as RNA molecules, of a defined size or range of sizes. The invention provides compositions, methods and kits for use in the production and preparation of small RNA molecules (including without limitation micro-RNA, siRNA, d-siRNA and e-siRNA) and other nucleic acids of various sizes.
摘要翻译： 本发明提供了制备具有规定尺寸或尺寸范围的核酸如RNA分子的方法。 本发明提供了用于生产和制备小RNA分子（包括但不限于微RNA，siRNA，d-siRNA和e-siRNA）和各种尺寸的其它核酸的组合物，方法和试剂盒。
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5.发明申请COMPOSITIONS AND METHODS FOR PREPARING SHORT RNA MOLECULES AND OTHER NUCLEIC ACIDS 审中-公开用于制备短链RNA分子和其他核酸的组合物和方法公开(公告)号：US20120108804A1
公开(公告)日：2012-05-03
申请号：US13190325
申请日：2011-07-25
申请人： Knut R. Madden , Adam N. Harris , Karl H. Hecker , Byung-in Lee
发明人： Knut R. Madden , Adam N. Harris , Karl H. Hecker , Byung-in Lee
IPC分类号： C07H21/02
CPC分类号： C07H21/02 , B01D15/3804 , C12N15/111 , C12N2330/31
摘要： The invention provides methods of preparing nucleic acids, such as RNA molecules, of a defined size or range of sizes. The invention provides compositions, methods and kits for use in the production and preparation of small RNA molecules (including without limitation micro-RNA, siRNA, d-siRNA and e-siRNA) and other nucleic acids of various sizes.
摘要翻译： 本发明提供了制备具有规定尺寸或尺寸范围的核酸如RNA分子的方法。 本发明提供了用于生产和制备小RNA分子（包括但不限于微RNA，siRNA，d-siRNA和e-siRNA）和各种尺寸的其它核酸的组合物，方法和试剂盒。
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