• 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

        Quantitative Measurement of Relative Gene Expression in Human Tumors

        互联网

        478
        Quantitative measurement of specific mRNA species is of major importance for approaching many fundamental questions in biology. Until now, quantitation of gene expression has usually been done by Northern blotting, but this procedure is relatively insensitive, requiring microgram amounts of RNA. Furthermore, unless linear ranges of RNA concentration are determined, the procedure is semiquantitative at best. Because of the limitations of Northern blotting, various strategies have been developed for quantitation of cDNA by polymerase chain reaction (PCR)-based methods (1 -3 ), most of them utilizing the principle of competitive PCR in which a synthetic segment is coamplified along the target DNA segment. We were interested in comparing the expression of drug target genes in very small samples of human tumor material, such as would be obtained from biopsies or even paraffin blocks. The competitive PCR was not entirely suitable for this purpose because: (1) the “input” RNA or DNA concentration must be known with precision in order to provide a normalization factor, and (2) our results suggested that the competitor and target were not necessarily amplified with the same efficiency even when the segments to be amplified were the same size (4 ). To overcome these problems, we developed an alternate method of PCR quantitation with the following key features:
        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