公开(公告)号：US20040191872A1

公开(公告)日：2004-09-30

申请号：US10438992

申请日：2003-05-16

Applicant: Korea Basic Science Institute

Inventor： Chaejoon Cheong ,  Chulhyun Lee ,  Hae-Kap Cheong ,  Jun-Soek Yoo ,  Eunha Hwang

IPC: C12P019/34 ,  C07H021/02

CPC classification number: C12P19/34

Abstract: A process for preparation of RNA using an affinity column and a DNAzyme is disclosed. By the process, a target RNA can be mass-produced with a high yield, a high resolution and a high purification efficiency. The process includes the steps of preparing a RNA adduct including a target RNA and a tail RNA sequence which is affixed to the 3null-terminal of the target RNA and can complementary combine with an oligo-dN sequence of an oligo-dN affinity column; purifying the RNA adduct by combining the RNA adduct to the oligo-dN affinity column; and obtaining the target RNA by cleaving the purified RNA adduct with a DNAzyme. Preferably, the process for preparation of RNA further includes the steps of removing a RNA fragment cleaved by the DNAzyme by using an affinity column; and decomposing the DNAzyme with a DNA nuclease, and then removing decomposed DNAzyme fragments with a gel filtration column.

Abstract translation: 公开了使用亲和柱和DNAzyme制备RNA的方法。 通过该方法，可以高产率，高分辨率和高纯化效率大量生产靶RNA。 该方法包括制备包含目标RNA和尾RNA序列的RNA加合物的步骤，所述RNA加合物附着于靶RNA的3'末端，并可与oligo-dN亲和柱的oligo-dN序列互补结合; 通过将RNA加合物与oligo-dN亲和柱结合来纯化RNA加合物; 并通过用DNA酶切割纯化的RNA加合物来获得靶RNA。 优选地，RNA的制备方法还包括通过使用亲和柱除去DNA酶切割的RNA片段的步骤; 并用DNA核酸酶分解DNA酶，然后用凝胶过滤柱除去分解的DNA酶片段。