Column for DNA separation by matched ion polynucleotide chromatography

    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

    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.

    Method and system for the preparation of cDNA

    公开(公告)号:US06521411B2

    公开(公告)日:2003-02-18

    申请号:US09808447

    申请日:2001-03-13

    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.

    Column for DNA separation by matched ion polynucleotide chromatography

    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

    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.