{"id":158,"date":"2015-11-03T00:00:00","date_gmt":"2015-11-03T00:00:00","guid":{"rendered":"https:\/\/www.thermofisher.com\/blog\/learning-at-the-bench\/qpcr-basic12\/"},"modified":"2025-04-17T15:12:59","modified_gmt":"2025-04-17T06:12:59","slug":"qpcr-basic12","status":"publish","type":"post","link":"https:\/\/www.thermofisher.com\/blog\/learning-at-the-bench\/qpcr-basic12\/","title":{"rendered":"\u5185\u5728\u6027\u30b3\u30f3\u30c8\u30ed\u30fc\u30eb\u306e\u9078\u629e\u3001\u8ff7\u3063\u305f\u3089\u3053\u308c\u3060\uff01\uff5c\u4eca\u3053\u305d\u672c\u6c17\u3067\u5fb9\u5e95\u7406\u89e3! \u30ea\u30a2\u30eb\u30bf\u30a4\u30e0PCR\u8b1b\u5ea7 \u7b2c12\u56de"},"content":{"rendered":"

\u4eca\u56de\u306f\u3001\u3069\u306e\u907a\u4f1d\u5b50\u3092\u5185\u5728\u6027\u30b3\u30f3\u30c8\u30ed\u30fc\u30eb\u907a\u4f1d\u5b50\u3068\u3057\u3066\u7528\u3044\u308c\u3070\u826f\u3044\u306e\u304b\u3001\u5185\u5728\u6027\u30b3\u30f3\u30c8\u30ed\u30fc\u30eb\u3092\u6700\u7d42\u7684\u306b\u6c7a\u5b9a\u3059\u308b\u305f\u3081\u306e\u3001\u691c\u8a0e\u306b\u5165\u308a\u307e\u3059\u3002<\/p>\n

\u904e\u53bb\u306b\u540c\u3058\u5b9f\u9a13\u7cfb\u3067\u5831\u544a\u304c\u3042\u308b\u5834\u5408<\/h2>\n

\u6700\u8fd1\u3067\u306f\u3001\u5185\u5728\u6027\u30b3\u30f3\u30c8\u30ed\u30fc\u30eb\u907a\u4f1d\u5b50\u3092\u691c\u8a0e\u3057\u305f\u5831\u544a\u304c\u5897\u3048\u3066\u304d\u3066\u3044\u307e\u3059\u3002\u3054\u81ea\u5206\u306e\u5b9f\u9a13\u7cfb\u306b\u3064\u3044\u3066\u4fe1\u983c\u3067\u304d\u308b\u904e\u53bb\u306e\u5831\u544a\u304c\u3042\u308b\u5834\u5408\u306f\u3001\u305d\u3053\u3067\u6700\u9069\u3068\u3055\u308c\u308b\u907a\u4f1d\u5b50\u3092\u7528\u3044\u308c\u3070\u3001\u307e\u305a\u554f\u984c\u306f\u7121\u3044\u3067\u3057\u3087\u3046\u3002<\/p>\n

\u904e\u53bb\u306b\u540c\u3058\u5b9f\u9a13\u7cfb\u3067\u5831\u544a\u304c\u7121\u3044\u5834\u5408<\/h2>\n

\u5f0a\u793e\u3067\u306f\u3001\u5185\u5728\u6027\u30b3\u30f3\u30c8\u30ed\u30fc\u30eb\u907a\u4f1d\u5b50\u3092\u691c\u8a0e\u3059\u308b\u305f\u3081\u306e\u30c4\u30fc\u30eb\u3001TaqMan\u00ae Array Human Endogenous Controls<\/a>\u3092\u63d0\u4f9b\u3057\u3066\u304a\u308a\u307e\u3059\u300296\u30a6\u30a7\u30ebPCR\u30d7\u30ec\u30fc\u30c8\u306b\u300132\u7a2e\u985e\u306e\u5185\u5728\u6027\u30b3\u30f3\u30c8\u30ed\u30fc\u30eb\u907a\u4f1d\u5b50\u3068\u306a\u308a\u3046\u308b\u907a\u4f1d\u5b50\u691c\u51fa\u7528TaqMan\u00ae\u30d7\u30ed\u30fc\u30d6\u3001\u30d7\u30e9\u30a4\u30de\u30fc\u30bb\u30c3\u30c8\u304c\u51cd\u7d50\u4e7e\u71e5\u3055\u308c\u305f\u72b6\u614b\u3067\u642d\u8f09\u3055\u308c\u3066\u3044\u307e\u3059\uff08\u56f31\uff09\u3002cDNA\u30b5\u30f3\u30d7\u30eb\u3068\u30de\u30b9\u30bf\u30fc\u30df\u30c3\u30af\u30b9\u3068\u89e3\u6790\u3057\u305f\u3044\u30b5\u30f3\u30d7\u30eb\u3092\u6df7\u305c\u3066\u52a0\u3048\u308b\u3060\u3051\u3067\u30013\u7a2e\u985e\u306e\u691c\u4f53\u306b\u95a2\u3057\u3066\u5185\u5728\u6027\u30b3\u30f3\u30c8\u30ed\u30fc\u30eb\u306e\u8a55\u4fa1\u304c\u53ef\u80fd\u3067\u3059\u3002<\/strong><\/p>\n

\"qpcr-basic12-fig1\"<\/a>

\u56f31 TaqMan\u00ae Human Endogenous Control Plate\u306b\u642d\u8f09\u3055\u308c\u3066\u3044\u308b\u5019\u88dc\u907a\u4f1d\u5b50\u300218S rRNA (18S)\u3001ribosomal protein large P0 (huP0)\u3001\u03b2-actin (hubA) \u3001peptidylprolyl isomerase A (cyclophilin A) (huCYC)\u3001GAPDH (huGAPDH)\u3001phosphoglycerate kinase 1 (huPGK)\u3001\u03b2-2-microglobulin (huB2m)\u3001\u03b2-glucuronidase (huGUS)\u3001hypoxanthine phosphoribosyltransferase 1 (huHPRT)\u3001TATA box binding protein (huTBP)\u3001transferrin receptor (p90, CD71) (huTfR)<\/p><\/div>\n

\u3067\u306f\u3055\u3063\u305d\u304f\u3001TaqMan\u00ae Array Human Endogenous Controls\u3092\u7528\u3044\u305f\u5185\u5728\u6027\u30b3\u30f3\u30c8\u30ed\u30fc\u30eb\u306e\u8a55\u4fa1\u3092\u307f\u3066\u304f\u3060\u3055\u3044\uff08\u56f32\uff09\u3002<\/p>\n

\u3053\u3053\u3067\u306f4\u3064\u306e\u30b5\u30f3\u30d7\u30eb\uff08\u9aa8\u9ac4\u3001\u813e\u81d3\u3001\u80f8\u817a\u3001\u80ba\u7531\u6765\u306e\u30c8\u30fc\u30bf\u30ebRNA\uff09\u3067\u3001\u5404\u907a\u4f1d\u5b50\u306e\u767a\u73fe\u5909\u52d5\u3092\u89e3\u6790\u3057\u3066\u3044\u307e\u3059\u300218S rRNA\u3001\u3042\u308b\u3044\u306f\u03b2-Actin\u304c\u30014\u30b5\u30f3\u30d7\u30eb\u9593\u3067\u306e\u767a\u73fe\u30ec\u30d9\u30eb\u306e\u5909\u52d5\u304c\u5c0f\u3055\u3044\u3053\u3068\u304c\u308f\u304b\u308a\u307e\u3059\u306d\u3002\u3053\u306e\u7d50\u679c\u304b\u3089\u300118S rRNA\u3001\u03b2-Actin\u304c\u5185\u5728\u6027\u30b3\u30f3\u30c8\u30ed\u30fc\u30eb\u907a\u4f1d\u5b50\u306b\u6700\u9069\u3067\u3042\u308a\u3001\u9006\u306bGAPDH\u3084transferrin receptor\u306f\u9069\u3057\u3066\u3044\u306a\u3044\u3068\u3044\u3048\u308b\u306e\u3067\u3059\u3002<\/strong><\/p>\n

\"qpcr-basic12-fig2\"<\/a>

\u56f32. Clontech\u793e\u304b\u3089\u8cfc\u5165\u3057\u305f\u9aa8\u9ac4\u3001\u813e\u81d3\u3001\u80f8\u817a\u3001\u80ba\u7531\u6765\u306e\u30c8\u30fc\u30bf\u30ebRNA\u3092MultiScribe\u2122 Reverse Transcriptase\u306b\u3088\u3063\u3066cDNA\u306b\u5909\u63db\u3002\u9aa8\u9ac4\u30b5\u30f3\u30d7\u30eb\u3092\u30ad\u30e3\u30ea\u30d6\u30ec\u30fc\u30bf\u30fc\u306b\u8a2d\u5b9a\u3057\u3001\u5404\u81d3\u5668\u9593\u3067\u306e\u907a\u4f1d\u5b50\u306e\u767a\u73fe\u91cf\u3092\u6bd4\u8f03\u3057\u305f\u3002\u7e26\u8ef8\u306fThreshold Cycle\u306e\u5dee\u3092\u793a\u3057\u3001\u4e0a\u306b\u4f38\u3073\u308b\u307b\u3069\u767a\u73fe\u91cf\u304c\u9ad8\u3044\u3053\u3068\u3092\u793a\u3059\u3002Threshold Cycle\u306e1\u30b5\u30a4\u30af\u30eb\u306e\u5dee\u306f\u3001\u907a\u4f1d\u5b50\u767a\u73fe\u91cf\u306b\u304a\u3044\u3066\u306f\u7d042\u500d\u306e\u5dee\u3092\u610f\u5473\u3059\u308b\u3002<\/p><\/div>\n

\u6700\u9069\u306a\u5185\u5728\u6027\u30b3\u30f3\u30c8\u30ed\u30fc\u30eb\u907a\u4f1d\u5b50\u5019\u88dc\u304c\u898b\u3064\u304b\u3063\u305f\u3089\u3001\u3042\u3068\u306f\u305d\u308c\u3092\u3054\u8cfc\u5165\u3044\u305f\u3060\u304f\u3060\u3051\u3067\u3001\u6761\u4ef6\u691c\u8a0e\u3092\u3057\u306a\u304f\u3068\u3082\u6b63\u78ba\u3067\u518d\u73fe\u6027\u306e\u9ad8\u3044\u7d50\u679c\u304c\u5f97\u3089\u308c\u307e\u3059\u3002
\n\n\t

\n\t\t18S rRNA\u3092\u9664\u304f\u307b\u3068\u3093\u3069\u306e\u30a2\u30c3\u30bb\u30a4\u3067\u306f\u30a8\u30af\u30bd\u30f3\u30b8\u30e3\u30f3\u30af\u30b7\u30e7\u30f3\u4e0a\u306bTaqMan\u00ae\u30d7\u30ed\u30fc\u30d6\u304c\u4f4d\u7f6e\u3057\u3066\u3044\u308b\u306e\u3067\u3001\u30b2\u30ce\u30e0DNA\u7531\u6765\u306e\u5897\u5e45\u691c\u51fa\u3092\u5fc3\u914d\u3059\u308b\u3053\u3068\u306a\u304f\u5b9f\u9a13\u3067\u304d\u307e\u3059\u3002\u307e\u305f\u3001\u30bf\u30fc\u30b2\u30c3\u30c8\u907a\u4f1d\u5b50\u3068\u5185\u5728\u6027\u30b3\u30f3\u30c8\u30ed\u30fc\u30eb\u907a\u4f1d\u5b50\u30921\u3064\u306e\u30c1\u30e5\u30fc\u30d6\u3067\u540c\u6642\u306b\u5897\u5e45\u3059\u308b\u3088\u3046\u30d7\u30e9\u30a4\u30de\u30fc\u6fc3\u5ea6\u304c\u6700\u9069\u5316\u3055\u308c\u3001multiplex\u306a\u53cd\u5fdc\u7cfb\u3067\u3082PCR\u304c\u6b63\u3057\u304f\u9032\u3080\u5185\u5728\u6027\u30ad\u30c3\u30c8\u3082\u3042\u308a\u307e\u3059\u3002\u901a\u5e38\u306f\u56f0\u96e3\u306amultiplex PCR\u304c\u624b\u8efd\u306b\u884c\u3048\u3001\u8cb4\u91cd\u306acDNA\u30b5\u30f3\u30d7\u30eb\u3092\u6709\u52b9\u306b\u5229\u7528\u3067\u304d\u307e\u3059\u3002\t<\/blockquote>\n\n\t

<\/p>\n\t\n\t
\n\u5f0a\u793e\u306e\u5185\u5728\u6027\u30b3\u30f3\u30c8\u30ed\u30fc\u30eb\u8a66\u85ac\u306f\u3001\u65e2\u306b\u69d8\u3005\u306a\u8ad6\u6587\u3067\u5831\u544a\u3055\u308c\u3066\u3044\u308b\u306e\u3067\u5b89\u5fc3\u3057\u3066\u3054\u4f7f\u7528\u3044\u305f\u3060\u3051\u307e\u3059\u3002<\/strong><\/p>\n

\u3053\u308c\u304b\u3089\u907a\u4f1d\u5b50\u306e\u767a\u73fe\u5b9a\u91cf\u89e3\u6790\u3092\u59cb\u3081\u3088\u3046\u3068\u3044\u3046\u65b9\u3001\u3042\u308b\u3044\u306f\u5b9f\u9a13\u4e2d\u306e\u65b9\u3082\u3001\u5185\u5728\u6027\u30b3\u30f3\u30c8\u30ed\u30fc\u30eb\u907a\u4f1d\u5b50\u306f\u5341\u5206\u306b\u691c\u8a0e\u3057\u3066\u9078\u629e\u3057\u3001\u7814\u7a76\u3092\u9032\u3081\u308b\u3053\u3068\u3092\u304a\u52e7\u3081\u3044\u305f\u3057\u307e\u3059\u3002<\/p>\n

\u30ea\u30a2\u30eb\u30bf\u30a4\u30e0PCR\u30cf\u30f3\u30c9\u30d6\u30c3\u30af \u7121\u6599\u30c0\u30a6\u30f3\u30ed\u30fc\u30c9<\/h2>\n

\u3053\u306e\u30cf\u30f3\u30c9\u30d6\u30c3\u30af\u3067\u306f\u3001\u30ea\u30a2\u30eb\u30bf\u30a4\u30e0PCR\u306e\u7406\u8ad6\u3084\u5b9f\u9a13\u30c7\u30b6\u30a4\u30f3\u306e\u8a2d\u8a08\u306a\u3069\u3001\u30ea\u30a2\u30eb\u30bf\u30a4\u30e0PCR\u306e\u57fa\u790e\u77e5\u8b58\u304c\u63b2\u8f09\u3055\u308c\u3066\u3044\u307e\u3059\u3002\u30ea\u30a2\u30eb\u30bf\u30a4\u30e0PCR\u3092\u59cb\u3081\u305f\u3070\u304b\u308a\u306e\u65b9\u3084\u3053\u308c\u304b\u3089\u5b9f\u9a13\u3092\u8003\u3048\u3066\u3044\u308b\u65b9\u306b\u3046\u3063\u3066\u3064\u3051\u306e\u30cf\u30f3\u30c9\u30d6\u30c3\u30af\u3067\u3059\u3002PDF\u30d5\u30a1\u30a4\u30eb\u306e\u30c0\u30a6\u30f3\u30ed\u30fc\u30c9\u3092\u3054\u5e0c\u671b\u306e\u65b9\u306f\u3001\u4e0b\u8a18\u30dc\u30bf\u30f3\u3088\u308a\u304a\u7533\u3057\u8fbc\u307f\u304f\u3060\u3055\u3044\u3002<\/p>\n

\u30c0\u30a6\u30f3\u30ed\u30fc\u30c9\u3059\u308b<\/a><\/p>\n

\u30ea\u30a2\u30eb\u30bf\u30a4\u30e0PCR\u30c8\u30e9\u30d6\u30eb\u30b7\u30e5\u30fc\u30c6\u30a3\u30f3\u30b0 \u7121\u6599\u30c0\u30a6\u30f3\u30ed\u30fc\u30c9<\/h2>\n

\u7570\u5e38\u306aS\u5b57\u72b6\u306e\u5897\u5e45\u66f2\u7dda\u3001NTC\u3067\u306e\u5897\u5e45\u691c\u51fa\u3001\u5897\u5e45\u304c\u898b\u3089\u308c\u306a\u3044\u306a\u3069\u3001\u30ea\u30a2\u30eb\u30bf\u30a4\u30e0PCR\u306e\u5b9f\u9a13\u4e2d\u306b\u56f0\u3063\u305f\u969b\u306e\u624b\u52a9\u3051\u306b\u306a\u308b\u60c5\u5831\u3092\u7d39\u4ecb\u3057\u3066\u3044\u307e\u3059\u3002PDF\u30d5\u30a1\u30a4\u30eb\u306e\u30c0\u30a6\u30f3\u30ed\u30fc\u30c9\u3092\u3054\u5e0c\u671b\u306e\u65b9\u306f\u3001\u4e0b\u8a18\u30dc\u30bf\u30f3\u3088\u308a\u304a\u7533\u3057\u8fbc\u307f\u304f\u3060\u3055\u3044\u3002<\/p>\n

\u30c0\u30a6\u30f3\u30ed\u30fc\u30c9\u3059\u308b<\/a><\/p>\n

 <\/p>\n

\u7814\u7a76\u7528\u306b\u306e\u307f\u4f7f\u7528\u3067\u304d\u307e\u3059\u3002\u8a3a\u65ad\u7528\u306b\u306f\u4f7f\u7528\u3044\u305f\u3060\u3051\u307e\u305b\u3093\u3002<\/p>\n","protected":false},"excerpt":{"rendered":"

\u5185\u5728\u6027\u30b3\u30f3\u30c8\u30ed\u30fc\u30eb\u907a\u4f1d\u5b50\u3092\u691c\u8a0e\u3059\u308b\u305f\u3081\u30c4\u30fc\u30eb\u3001TaqMan\u00ae Array Human Endogenous 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