{"id":294,"date":"2020-12-02T12:30:59","date_gmt":"2020-12-02T03:30:59","guid":{"rendered":"https:\/\/www.promega.co.jp\/resources\/faq\/?p=294"},"modified":"2020-12-15T14:41:51","modified_gmt":"2020-12-15T05:41:51","slug":"q98","status":"publish","type":"post","link":"https:\/\/www.promega.co.jp\/resources\/faq\/amplification-quantification\/pcr-and-rt\/rt-pcr\/rt-pcr-basic-info\/q98","title":{"rendered":"RT-PCR\u306b\u304a\u3044\u3066RNA\u306e2\u6b21\u69cb\u9020\u306b\u95a2\u3059\u308b\u554f\u984c\u3092\u89e3\u6c7a\u3059\u308b\u305f\u3081\u306b\u6709\u52b9\u306a\u65b9\u6cd5\u3068\u306f\u4f55\u3067\u3059\u304b\u3002"},"content":{"rendered":"\n

AMV RT\u306fM-MLV RT\u3088\u308a\u3082\u9ad8\u3044\u53cd\u5fdc\u6e29\u5ea6\u3067\u6d3b\u6027\u3092\u793a\u3057\u307e\u3059\u3002\u3053\u306e\u305f\u3081AMV RT\u306f\u5f37\u56fa\u306a2\u6b21\u69cb\u9020\u3092\u6301\u3064\u92f3\u578b\u306b\u5bfe\u3057\u3066\u306f\u3088\u308a\u826f\u3044\u9078\u629e\u3067\u3042\u308b\u3068\u8003\u3048\u3089\u308c\u307e\u3059\u3002AMV RT\u306f58\u2103\u307e\u3067\u6d3b\u6027\u3092\u7dad\u6301\u3057\u307e\u3059\u3002\u3057\u304b\u3057\u306a\u304c\u3089\u300150\u2103\u4ee5\u4e0a\u3067\u306f\u3059\u3050\u306b\u6d3b\u6027\u306f\u4f4e\u4e0b\u3057\u3001\u9577\u6642\u9593\u306e\u6d3b\u6027\u7dad\u6301\u306f\u3067\u304d\u307e\u305b\u3093\u3002\u9ad8\u6e29\u3067\u306e\u9006\u8ee2\u5199\u3092\u884c\u3046\u6642\u306b\u8003\u616e\u3059\u308b\u3053\u3068\u3068\u3057\u3066\u3001\u30d7\u30e9\u30a4\u30de\u30fc\u306e\u30a2\u30cb\u30fc\u30ea\u30f3\u30b0\u6e29\u5ea6\u304c\u6319\u3052\u3089\u308c\u307e\u3059\u3002\u3088\u308a\u9ad8\u6e29\u3067\u306e\u53cd\u5fdc\u3067\u306f42\u2103\u3092\u6700\u9069\u306e\u30a2\u30cb\u30fc\u30ea\u30f3\u30b0\u6e29\u5ea6\u3068\u3059\u308boligo(dT)\u30d7\u30e9\u30a4\u30de\u30fc\u3092\u4f7f\u3048\u306a\u3044\u53ef\u80fd\u6027\u304c\u3042\u308a\u307e\u3059\u3002\u3053\u306e\u3088\u3046\u306b\u9ad8\u3044\u53cd\u5fdc\u6e29\u5ea6\u4e0b\u3067\u306f\u3001\u76ee\u7684\u306e\u907a\u4f1d\u5b50\u306b\u7279\u7570\u7684\u306a\u30d7\u30e9\u30a4\u30de\u30fc\u3092\u4f7f\u3046\u3053\u3068\u304c\u671b\u307e\u3057\u3044\u3067\u3059\u3002<\/p>\n\n\n\n

RNA\u306e\u69cb\u9020\u304c\u554f\u984c\u3068\u306a\u308b\u30b5\u30f3\u30d7\u30eb\u3092\u6271\u3046\u5834\u5408\u306b\u7528\u3044\u308b\u3079\u304d\u305d\u306e\u4ed6\u306e\u65b9\u6cd5\u3068\u3057\u30662\uff5e5% DMSO\u306e\u6dfb\u52a0\u304c\u3042\u3052\u3089\u308c\u307e\u3059\u3002DMSO\u306f\u5869\u57fa\u5bfe\u3092\u89e3\u96e2\u3055\u305b\u308b\u305f\u3081\u3001\u5897\u5e45\u91cf\u3092\u5897\u52a0\u3055\u305b\u308b\u3053\u3068\u304c\u3067\u304d\u307e\u3059(1)\u3002 Sodium pyrophosphate (<=4mM)\u3082\u3001RNA\u306e2\u6b21\u69cb\u9020\u3084\u7b2c1\u9396 cDNA\u306e\u53ce\u91cf\u3092\u5897\u52a0\u3059\u308b\u305f\u3081\u306e\u624b\u6bb5\u3068\u3057\u3066\u3001AMV RT\u3092\u7528\u3044\u305f2\u6bb5\u968e\u306eRT-PCR\u306eRT\u53cd\u5fdc\u306b\u6dfb\u52a0\u3055\u308c\u307e\u3059(2)\u3002pyrophosphate\u306f\u3001\u9175\u7d20\u306eDNA\u4f38\u9577\u53cd\u5fdc\u3092\u9045\u304f\u3055\u305b\u3001\u5f37\u56fa\u306aRNA\u306e2\u6b21\u69cb\u9020\u306b\u3088\u308b\u92f3\u578b\u304b\u3089\u306e\u9175\u7d20\u306e\u843d\u4e0b\u3092\u8d77\u3053\u308a\u306b\u304f\u304f\u3057\u307e\u3059\u3002M-MLV RT\u3067\u306f\u3001pyrophosphate\u3078\u306e\u611f\u53d7\u6027\u304c\u5897\u52a0\u3055\u308c\u3066\u3044\u308b\u305f\u3081\u3001\u4f7f\u7528\u3057\u306a\u3044\u3067\u304f\u3060\u3055\u3044(3)\u3002\uff08PN71-Q&A)<\/p>\n\n\n\n

\u53c2\u8003\u6587\u732e\uff1a<\/strong><\/small><\/p>\n\n\n\n

  1. Frackman, S. et al. (1998) Promega Notes 65, 27.<\/li>
  2. Kacian, D.L. and Myers, J.C. (1976) Proc. Natl. Acad. Sci. USA 73, 2191.<\/li>
  3. Roth, M.J., Tanese, N. and Goff, S.P. (1985) J. Biol. Chem. 260, 9326.<\/li><\/ol>\n","protected":false},"excerpt":{"rendered":"

    AMV RT\u306fM-MLV RT\u3088\u308a\u3082\u9ad8\u3044\u53cd\u5fdc\u6e29\u5ea6\u3067\u6d3b\u6027\u3092\u793a\u3057\u307e\u3059\u3002\u3053\u306e\u305f\u3081AMV RT\u306f\u5f37\u56fa\u306a2\u6b21\u69cb\u9020\u3092\u6301\u3064\u92f3\u578b\u306b\u5bfe\u3057\u3066\u306f\u3088\u308a\u826f\u3044\u9078\u629e\u3067\u3042\u308b\u3068\u8003\u3048\u3089\u308c\u307e\u3059\u3002AMV RT\u306f58\u2103\u307e\u3067\u6d3b\u6027\u3092\u7dad\u6301\u3057\u307e\u3059\u3002\u3057\u304b\u3057\u306a\u304c\u3089\u300150\u2103\u4ee5 … <\/p>\n","protected":false},"author":16,"featured_media":0,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":[],"categories":[94],"tags":[],"_links":{"self":[{"href":"https:\/\/www.promega.co.jp\/resources\/faq\/wp-json\/wp\/v2\/posts\/294"}],"collection":[{"href":"https:\/\/www.promega.co.jp\/resources\/faq\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/www.promega.co.jp\/resources\/faq\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/www.promega.co.jp\/resources\/faq\/wp-json\/wp\/v2\/users\/16"}],"replies":[{"embeddable":true,"href":"https:\/\/www.promega.co.jp\/resources\/faq\/wp-json\/wp\/v2\/comments?post=294"}],"version-history":[{"count":1,"href":"https:\/\/www.promega.co.jp\/resources\/faq\/wp-json\/wp\/v2\/posts\/294\/revisions"}],"predecessor-version":[{"id":295,"href":"https:\/\/www.promega.co.jp\/resources\/faq\/wp-json\/wp\/v2\/posts\/294\/revisions\/295"}],"wp:attachment":[{"href":"https:\/\/www.promega.co.jp\/resources\/faq\/wp-json\/wp\/v2\/media?parent=294"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/www.promega.co.jp\/resources\/faq\/wp-json\/wp\/v2\/categories?post=294"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/www.promega.co.jp\/resources\/faq\/wp-json\/wp\/v2\/tags?post=294"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}