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Method of identification and cloning differentially expressed messenger RNAs

机译:鉴定和克隆差异表达信使RNA的方法

摘要

A method of identification of differentially expressed messenger RNA (mRNA) which consists of synthesizing from a set of sequences of mRNA sets of fragments of complementary DNA (cDNA), which are separated with the aid of gel electrophoresis and the pictures of separation of the cDNA from different types of cells are compared and fragments with the aid of restriction nucleases. A method of cloning of differentially expressed mRNAs consists of synthesizing from sets of sequences of MRNAs from different types of cells sets of fragments of complementary DNA (cDNA) which are separated with the aid of gel electrophoresis, the pictures of the separation of the cDNA from different types of cells are compared, fragments of cDNA with different signal intensities are separated from the gel, amplified with the aid of a polymerase chain reaction and clones to a plasmid or phage vector. For the formation of the set of fragments one carries out cleavage of the cDNA with the aid of restriction endonucleases and uses only those fragments of cDNA that correspond to the 3' or 5' end regions of the mRNAs.
机译:一种鉴定差异表达信使RNA(mRNA)的方法,该方法包括从一组mRNA序列中合成互补DNA(cDNA)的片段集,这些片段借助凝胶电泳和cDNA分离图进行分离比较来自不同类型细胞的细胞,并借助限制性核酸酶切割片段。克隆差异表达mRNA的方法包括:从不同类型细胞的MRNA序列集中合成互补DNA(cDNA)片段集,这些片段借助凝胶电泳分离,从中分离cDNA的图片比较不同类型的细胞,将具有不同信号强度的cDNA片段从凝胶中分离出来,借助聚合酶链反应进行扩增,然后克隆到质粒或噬菌体载体中。为了形成片段组,人们借助于限制性核酸内切酶进行cDNA的切割,并且仅使用与mRNA的3'或5'末端区域相对应的那些cDNA片段。

著录项

  • 公开/公告号US5814445A

    专利类型

  • 公开/公告日1998-09-29

    原文格式PDF

  • 申请/专利权人 NEW YORK BLOOD CENTER;

    申请/专利号US19950499899

  • 发明设计人 NATALIA B. IVANOVA;ALEXANDER V. BELYAVSKY;

    申请日1995-07-11

  • 分类号C12Q1/68;C12P19/34;C07H21/04;

  • 国家 US

  • 入库时间 2022-08-22 02:38:32

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