• 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

        Qualitative and Quantitative DNA and RNA Analysis by Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry

        互联网

        621
        Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) gives extremely precise reading of mass-to-charge ratios (two analytes differ by 1 Da can be distinguished) and provides high sensitivity (less than 1 fmole of a DNA oligonucleotide can be detected), allowing DNA quantifications with single base specificity and single DNA molecule sensitivity (coupled with polymerase chain reaction [PCR]). To quantify a DNA sequence of interest, a competitive synthetic (60-80 bases) oligonucleotide standard with an artificial single base mutation in the middle is introduced, and these two virtually identical sequences are co-amplified by PCR. A third primer (extension primer) is designed to anneal to the region immediately upstream of the mutation site. Depending on the specific mutation introduced and the ddNTP/dNTP mixtures used, either one or two bases are added to the extension primer to produce two extension products from the two templates. Last, the two extension products are detected and quantified by high-throughput MALDI-TOF MS. In addition, with an improved primer extension method called single allele base extension reaction (SABER), rare mutant DNA can be robustly detected even when normal DNA is present at 50-fold or more than the DNA mutants.
        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