• 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

        Generation of Longer cDNA Fragments from SAGE Tags for Gene Identification

        互联网

        440
        Serial analysis of gene expression (SAGE) is a powerful technique for genomewide analysis of gene expression (1 14 ). However, almost two-thirds of SAGE tags cannot be used directly for gene identification for two reasons. First, many of SAGE tags match to multiple known expressed sequences because of the short length of SAGE tag sequences (12 14 ). Second, many SAGE tags do not match any known expressed sequences because the sequences corresponding to these SAGE tags have not been identified yet (2 14 ). These problems substantially diminish the power of the SAGE technique. The GLGI (Generation of Longer complementary DNA [cDNA] fragments from SAGE tags for Gene Identification) technique was designed to solve these two problems (15 ). The basic principle of GLGI is to use the SAGE tag as the sense primer, and anchored oligo-(dT) primers as the antisense primer to amplify the original 3′ cDNA from which the SAGE tag was derived. The size of 3′ cDNA will be hundreds of bases longer, which is long enough for solving the two problems. In a typical SAGE project, hundreds or thousands of SAGE tags need to be further analyzed. To facilitate such large-scale performance, we developed the GLGI method into a high-throughput procedure (16 ). In this high-throughput GLGI procedure, 3′ cDNAs starting from the last CATG are used as the templates for GLGI amplification, a SAGE tag sequence is used as the sense primer, and a universal sequence located at the 3′ end of all the cDNA templates generated from anchored oligo(dT) primers is used as antisense primer to amplify the original 3′ cDNA template from which the SAGE tag was derived.
        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