• 我要登录|
  • 免费注册
    |
  • 我的丁香通
    • 企业机构:
    • 成为企业机构
    • 个人用户:
    • 个人中心
  • 移动端
    移动端
丁香通 logo丁香实验_LOGO
搜实验

    大家都在搜

      大家都在搜

        0 人通过求购买到了急需的产品
        免费发布求购
        发布求购
        点赞
        收藏
        wx-share
        分享

        Primer Extension Assay

        互联网

        1709
        A number of protocols are available for the analysis of RNA transcripts: Northern analysis (see Chapter 18 ), S1 mapping, primer extension, RNase AT1 mapping (see Chapter 20 ), and the more recent reverse transcriptase/PCR amplification (see Chapters 22 and24 ). These protocols have been applied to the analysis of plant RNAs and all vary in the degree of sensitivity of detection and the information generated. Primer extension assays have been used mainly in mapping of 5′ termini of RNA transcripts, but have also been of value, in, for example, analysis of nuclear pre-mRNA splicing (1 3 ), and in the detection of low abundance RNA species (2 ). There are four steps involved in performing a primer extension assay. First, selection and preparation of a labeled primer complementary to the RNA transcript of interest. Second, hybridization of the primer complementary to a region of the RNA under study. Third, extension from the primer that is catalyzed by an RNA-dependent DNA polymerase (reverse transcriptase) using RNA as the template to synthesize a cDNA strand. Fourth, analysis of the extended cDNA products on denaturing polyacrylaminde gels and autoradiography.
        ad image
        提问
        扫一扫
        丁香实验小程序二维码
        实验小助手
        丁香实验公众号二维码
        扫码领资料
        反馈
        TOP
        打开小程序