{"id":6507,"date":"2020-05-18T15:22:08","date_gmt":"2020-05-18T06:22:08","guid":{"rendered":"http:\/\/admin.acceleratingscience.com\/learning-at-the-bench\/?p=6507"},"modified":"2025-04-15T16:21:49","modified_gmt":"2025-04-15T07:21:49","slug":"sybr_qpcr_gsd_ts_1","status":"publish","type":"post","link":"https:\/\/www.thermofisher.com\/blog\/learning-at-the-bench\/sybr_qpcr_gsd_ts_1\/","title":{"rendered":"\u304a\u60a9\u307f\u89e3\u6c7a\uff01SYBR Green\u53cd\u5fdc\u7cfb\u306e\u6700\u9069\u5316"},"content":{"rendered":"

\u30ea\u30a2\u30eb\u30bf\u30a4\u30e0PCR\u306bSYBR\u2122 Green\u3092\u4f7f\u308f\u308c\u3066\u3044\u308b\u65b9\u3067\u3001\u3053\u3093\u306a\u304a\u60a9\u307f\u3092\u6301\u3063\u3066\u3044\u308b\u65b9\u306f\u610f\u5916\u3068\u591a\u3044\u306e\u3067\u306f\u306a\u3044\u3067\u3057\u3087\u3046\u304b\u3002
\n\u300c\u975e\u7279\u7570\u7684\u306a\u5897\u5e45\u304c\u8d77\u304d\u3066\u3057\u307e\u3063\u3066\u3044\u308b\u300d
\n\u300c\u53cd\u5fdc\u6761\u4ef6\u304c\u60aa\u3044\u306e\u3067\u306f\u3068\u611f\u3058\u308b\u304c\u3001\u3069\u3046\u6761\u4ef6\u3092\u5909\u3048\u308c\u3070\u3044\u3044\u306e\u304b\u308f\u304b\u3089\u306a\u3044\u300d
\n\u305d\u3053\u3067\u4eca\u56de\u306f\u3001\u304a\u60a9\u307f\u3092\u89e3\u6c7a\u3059\u308bSYBR Green\u53cd\u5fdc\u7cfb\u306e\u6761\u4ef6\u6700\u9069\u5316\u306b\u3064\u3044\u3066\u3001\u3054\u7d39\u4ecb\u3057\u307e\u3059\u3002<\/p>\n

\u7406\u60f3\u7684\u306a\u53cd\u5fdc\u6761\u4ef6\u306b\u306a\u3063\u3066\u3044\u308b\u304b\u306e\u78ba\u8a8d\u65b9\u6cd5<\/h2>\n

SYBR Green\u306e\u53cd\u5fdc\u7cfb\u306e\u5834\u5408\u306f\u300c\u975e\u7279\u7570\u7684\u306a\u53cd\u5fdc\u304c\u8d77\u304d\u3066\u3044\u306a\u3044\u304b\uff1f\u300d\u3092\u78ba\u8a8d\u3059\u308b\u5fc5\u8981\u304c\u3042\u308a\u307e\u3059\u3002
\nSYBR Green\u306f\u53cd\u5fdc\u6db2\u4e2d\u306b\u5b58\u5728\u3059\u308b\u3059\u3079\u3066\u306e2\u672c\u9396DNA\u3092\u691c\u51fa\u3059\u308b\u305f\u3081\u3001\u975e\u7279\u7570\u7684\u306a\u5897\u5e45\u3082\u30bf\u30fc\u30b2\u30c3\u30c8\u306e\u5897\u5e45\u3082\u540c\u3058\u5897\u5e45\u3068\u3057\u3066\u691c\u51fa\u3055\u308c\u3001\u533a\u5225\u304c\u3064\u304b\u306a\u3044\u304b\u3089\u3067\u3059\u3002\u305d\u3082\u305d\u3082SYBR Green\u306f2\u672c\u9396DNA\u306e\u9593\u306b\u5165\u308a\u8fbc\u3093\u3067\uff08\u30a4\u30f3\u30bf\u30fc\u30ab\u30ec\u30fc\u30c8\u3057\u3066\uff09\u86cd\u5149\u3092\u767a\u3059\u308b\u8272\u7d20\u3067\u3059\u3002\u305d\u306e\u305f\u3081\u3001\u4eee\u306b\u30d7\u30e9\u30a4\u30de\u30fc\u540c\u58eb\u304c\u7d50\u5408\u3057\u3066\u751f\u6210\u3055\u308c\u3066\u3057\u307e\u3063\u305f\u30d7\u30e9\u30a4\u30de\u30fc\u30c0\u30a4\u30de\u30fc\u3067\u3042\u3063\u3066\u3082\u86cd\u5149\u304c\u691c\u51fa\u3055\u308c\u307e\u3059\u3002\u76ee\u7684\u306e\u5897\u5e45\u7523\u7269\u4ee5\u5916\u3082\u691c\u51fa\u3059\u308b\u305f\u3081\u3001\u3082\u3057\u4eee\u306b\u975e\u7279\u7570\u7684\u306a\u5897\u5e45\u304c\u3042\u3063\u305f\u5834\u5408\u3001\u3053\u3061\u3089\u3082PCR\u306e\u5897\u5e45\u3068\u8aa4\u3063\u3066\u6349\u3048\u3066\u3057\u307e\u3044\u307e\u3059\u3002
\n\u76ee\u7684\u4ee5\u5916\u306e\u5897\u5e45\u304c\u8d77\u304d\u3066\u3044\u306a\u3044\u304b\u3092\u78ba\u8a8d\u3059\u308b\u305f\u3081\u306b\u306f\u89e3\u96e2\u66f2\u7dda\uff08Dissociation curve\u3084Melt curve\uff09\u3092\u5f15\u304d\u307e\u3059\u3002\u3053\u308c\u306f\u30ea\u30a2\u30eb\u30bf\u30a4\u30e0PCR\u88c5\u7f6e\u306b\u30d7\u30ed\u30b0\u30e9\u30e0\u3068\u3057\u3066\u7d44\u307f\u8fbc\u307e\u308c\u3066\u304a\u308a\u3001PCR\u53cd\u5fdc\u306e\u5f8c\u306b\u7d9a\u3051\u3066\u5b9f\u65bd\u3057\u307e\u3059\u3002\u4ed5\u7d44\u307f\u306f\u7c21\u5358\u3067\u3059\u3002\u3086\u3063\u304f\u308a\u3068\u6e29\u5ea6\u3092\u4e0a\u3052\u3064\u3064\u3001\u86cd\u5149\u5024\u306e\u30c7\u30fc\u30bf\u3092\u56de\u53ce\u3059\u308b\u3060\u3051\u3067\u3059\u3002\u6e29\u5ea6\u3092\u4e0a\u3052\u3066\u3044\u304f\u3068\u3001\u3042\u308b\u6e29\u5ea6\u306b\u5230\u9054\u3057\u305f\u3068\u304d\u306b\u3001\u305d\u308c\u307e\u30672\u672c\u9396\u3067\u3042\u3063\u305fDNA\u304c1\u672c\u9396\u306b\u89e3\u96e2\u3057\u307e\u3059\u3002\u3053\u306e\u6e29\u5ea6\u304cTm\u5024\u3067\u3059\u3002Tm\u5024\u306f\u5897\u5e45\u7523\u7269\u306e\u9577\u3055\u3084GC\u542b\u91cf\u3068\u3044\u3063\u305f\u914d\u5217\u306b\u3088\u3063\u3066\u5909\u308f\u308a\u307e\u3059\u3002\u305d\u306e\u305f\u3081\u5225\u306e\u5897\u5e45\u7523\u7269\u304c\u3042\u308b\u5834\u5408\u3001Tm\u5024\u304c\u7570\u306a\u308a\u307e\u3059\u306e\u3067\u3001\u86cd\u5149\u5024\u306e\u5909\u5316\u304c2\u56de\u307f\u3089\u308c\u308b\u3053\u3068\u306b\u306a\u308a\u307e\u3059\u3002
\n\u3053\u306e\u86cd\u5149\u5024\u306e\u5909\u5316\u3092\u898b\u3084\u3059\u304f\u3059\u308b\u305f\u3081\u306b\u86cd\u5149\u5024\u3092\u5fae\u5206\u3057\u305f\u306e\u304c\u3001\u56f31\u306e\u53f3\u4e0b\u306b\u3042\u308b\u30b0\u30e9\u30d5\u3067\u3059\u3002\u30d4\u30fc\u30af\u306f\u86cd\u5149\u5024\u306e\u5287\u7684\u306a\u5909\u5316\u304c\u8d77\u304d\u305f\u6e29\u5ea6\uff08\uff1dTm\u5024\uff09\u306b\u51fa\u307e\u3059\u3002<\/p>\n

\"\"\u56f31 \u89e3\u96e2\u66f2\u7dda\u306e\u63cf\u753b\u306e\u539f\u7406\u3068\u6982\u5ff5\u56f3<\/p>\n

\u30d4\u30fc\u30af\u304c1\u3064\u3067\u3042\u308c\u3070\u5897\u5e45\u7523\u7269\u306f1\u3064\u3001\u30d4\u30fc\u30af\u304c\u8907\u6570\u3042\u308b\u3088\u3046\u3067\u3042\u308c\u3070\u5897\u5e45\u7523\u7269\u304c\u8907\u6570\u3042\u308b\u3068\u3044\u3048\u307e\u3059\u3002\u56f32\u306f\u8907\u6570\u306e\u30d4\u30fc\u30af\u304c\u898b\u3089\u308c\u308b\u4f8b\u3067\u3059\u3002\u3082\u3057\u3053\u306e\u3088\u3046\u306b\u8907\u6570\u306e\u30d4\u30fc\u30af\u304c\u3042\u308b\u3088\u3046\u3067\u3042\u308c\u3070\u6761\u4ef6\u306e\u6700\u9069\u5316\u304c\u5fc5\u8981\u3068\u306a\u308a\u307e\u3059\u3002<\/p>\n

\"\"\u56f32 \u8907\u6570\u306e\u30d4\u30fc\u30af\u304c\u307f\u3089\u308c\u308b\u4f8b<\/p>\n

\u30d7\u30e9\u30a4\u30de\u30fc\u6fc3\u5ea6\u306e\u6700\u9069\u5316<\/h2>\n

\u3044\u3088\u3044\u3088\u53cd\u5fdc\u6761\u4ef6\u306e\u6700\u9069\u5316\u306e\u8a71\u3067\u3059\u3002
\n1\u3064\u76ee\u306e\u65b9\u6cd5\u306f\u30d7\u30e9\u30a4\u30de\u30fc\u6fc3\u5ea6\u3092\u5909\u3048\u308b\u3053\u3068\u3067\u3059\u3002\u4e00\u822c\u7684\u306b\u30d7\u30e9\u30a4\u30de\u30fc\u6fc3\u5ea6\u304c\u9ad8\u3044\u65b9\u304c\u975e\u7279\u7570\u7684\u306a\u5897\u5e45\u3082\u8d77\u3053\u308a\u3084\u3059\u3044\u3067\u3059\u3002\u975e\u7279\u7570\u7684\u306a\u5897\u5e45\u304c\u307f\u3089\u308c\u3066\u3044\u308b\u5834\u5408\u306f\u3001\u30d7\u30e9\u30a4\u30de\u30fc\u6fc3\u5ea6\u3092\u4e0b\u3052\u308b\u3068\u6539\u5584\u3059\u308b\u5834\u5408\u304c\u3042\u308a\u307e\u3059\u3002\u305f\u3068\u3048\u3070\u3001Applied Biosystems\u2122 PowerUp SYBR Green Master Mix\u306f\u3001300\u301c800 nM\u306e\u7bc4\u56f2\u306e\u30d7\u30e9\u30a4\u30de\u30fc\u6fc3\u5ea6\u3067\u6700\u9069\u306b\u6a5f\u80fd\u3057\u307e\u3059\u3002\u4e00\u822c\u7684\u306b\u306f\u6fc3\u5ea6\u304c\u6fc3\u3044\u65b9\u304cPCR\u52b9\u7387\u304c\u9ad8\u304f\u306a\u308a\u307e\u3059\u306e\u3067\u3001\u6700\u521d\u306f\u6fc3\u3044\u6761\u4ef6\u3067\u8a66\u3057\u3066\u3001\u4e0a\u624b\u304f\u3044\u304b\u306a\u3044\u5834\u5408\u306b\u8584\u3044\u6761\u4ef6\u3067\u8a66\u3059\u306e\u304c\u30aa\u30b9\u30b9\u30e1\u3067\u3059\u3002
\n\u307e\u305f\u5fc5\u305a\u3057\u3082forward\u3068reverse\u306e\u6fc3\u5ea6\u304c\u4e00\u81f4\u3057\u3066\u3044\u308b\u5fc5\u8981\u306f\u3042\u308a\u307e\u305b\u3093\u3002\u5834\u5408\u306b\u3088\u3063\u3066\u306f\u3069\u3061\u3089\u304b\u306e\u30d7\u30e9\u30a4\u30de\u30fc\u306e\u307f\u6fc3\u5ea6\u3092\u5909\u66f4\u3059\u308b\u3053\u3068\u3067\u6539\u5584\u3059\u308b\u30b1\u30fc\u30b9\u3082\u3042\u308a\u307e\u3059\u3002\u30d7\u30e9\u30a4\u30de\u30fc\u6fc3\u5ea6\u306e\u9069\u5207\u306a\u7bc4\u56f2\u306f\u3001\u30de\u30b9\u30bf\u30fc\u30df\u30c3\u30af\u30b9\u306b\u3088\u3063\u3066\u6c7a\u307e\u308a\u307e\u3059\u3002\u88fd\u54c1\u60c5\u5831\u3092\u78ba\u8a8d\u306e\u4e0a\u3001\u6fc3\u5ea6\u3092\u5909\u3048\u3066\u304f\u3060\u3055\u3044\u3002<\/p>\n

\u53cd\u5fdc\u6761\u4ef6\u3000\u6e29\u5ea6\u306e\u6700\u9069\u5316<\/h2>\n

\u975e\u7279\u7570\u7684\u306a\u5897\u5e45\u306fAnnealing\uff0fExtension\u306e\u6e29\u5ea6\u3092\u4e0a\u3052\u308b\u3068\u6539\u5584\u3059\u308b\u50be\u5411\u304c\u3042\u308a\u307e\u3059\u3002\u4e00\u822c\u7684\u306aAnnealing\uff0fExtension\u306e\u6e29\u5ea6\u306f60\u2103\u3067\u3059\u304c\u3001\u3053\u308c\u30921\u2103\u523b\u307f\u306764\u2103\u7a0b\u5ea6\u307e\u3067\u4e0a\u3052\u3066\u975e\u7279\u7570\u7684\u306a\u5897\u5e45\u304c\u306a\u304f\u306a\u308b\u304b\u78ba\u8a8d\u3057\u307e\u3059\u3002<\/p>\n

\u305f\u3060\u3001\u6e29\u5ea6\u3092\u5909\u3048\u308b\u3068\u3044\u3046\u3053\u3068\u306f\u5225\u3005\u306b\u30e9\u30f3\u3057\u306a\u3051\u308c\u3070\u3044\u3051\u306a\u3044\u306e\u304b\u3068\u3001\u9762\u5012\u306b\u611f\u3058\u308b\u65b9\u3082\u3044\u3089\u3063\u3057\u3083\u308b\u3067\u3057\u3087\u3046\u304b\u3002\u3082\u3057\u304a\u4f7f\u3044\u306e\u88c5\u7f6e\u304cApplied Biosystems\u2122 StepOneplus\u2122\u30ea\u30a2\u30eb\u30bf\u30a4\u30e0PCR\u30b7\u30b9\u30c6\u30e0\u3084 Applied Biosystems\u2122 QunantStudio3\/5\u30ea\u30a2\u30eb\u30bf\u30a4\u30e0PCR\u30b7\u30b9\u30c6\u30e0\u306e\u3088\u3046\u306aVeriFlex\u3092\u642d\u8f09\u3057\u3066\u3044\u308b\u88c5\u7f6e\u3067\u3042\u308c\u3070\u72ec\u7acb\u3057\u305f\u6e29\u5ea6\u5236\u5fa1\u304c\u53ef\u80fd\u3067\u3059\u306e\u3067\u30011\u56de\u306e\u30e9\u30f3\u3067\u8907\u6570\u306eAnnealing\uff0fExtention\u6e29\u5ea6\u3092\u691c\u8a0e\u3067\u304d\u307e\u3059\u3002\u8a2d\u5b9a\u65b9\u6cd5\u306a\u3069\u306e\u8a73\u7d30\u3092\u3054\u5e0c\u671b\u3067\u3042\u308c\u3070\u30c6\u30af\u30cb\u30ab\u30eb\u30b5\u30dd\u30fc\u30c8<\/a>\u306b\u304a\u554f\u5408\u305b\u304f\u3060\u3055\u3044\u3002<\/p>\n

\u6700\u5f8c\u306e\u624b\u3000\u30d7\u30e9\u30a4\u30de\u30fc\u306e\u518d\u8a2d\u8a08<\/h2>\n

\u6fc3\u5ea6\u3092\u5909\u3048\u3066\u3082\u3001\u6e29\u5ea6\u3092\u5909\u3048\u3066\u3082\u6539\u5584\u3057\u306a\u3044\u3068\u3044\u3046\u5834\u5408\u306f\u30d7\u30e9\u30a4\u30de\u30fc\u3092\u518d\u8a2d\u8a08\u3057\u307e\u3059\u3002
\n\u30d7\u30e9\u30a4\u30de\u30fc\u30c0\u30a4\u30de\u30fc\u3092\u542b\u3080\u975e\u7279\u7570\u7684\u306a\u5897\u5e45\u304c\u8d77\u3053\u308b\u304b\u306f\u30d7\u30e9\u30a4\u30de\u30fc\u306e\u914d\u5217\u306b\u5de6\u53f3\u3055\u308c\u307e\u3059\u3002\u6fc3\u5ea6\u3084\u6e29\u5ea6\u3092\u8907\u6570\u6761\u4ef6\u8a66\u3057\u3066\u3082\u6539\u5584\u3057\u306a\u3044\u5834\u5408\u306f\u601d\u3044\u5207\u3063\u3066\u65b0\u3057\u3044\u30d7\u30e9\u30a4\u30de\u30fc\u3092\u8a2d\u8a08\u3059\u308b\u306e\u304c\u65e9\u3044\u3067\u3059\u3002\u3055\u307e\u3056\u307e\u306a\u6761\u4ef6\u3092\u8a66\u3059\u3088\u308a\u3082\u30d7\u30e9\u30a4\u30de\u30fc\u3092\u65b0\u3057\u304f\u3059\u308b\u65b9\u304c\u30b3\u30b9\u30c8\u9762\u3067\u3082\u304a\u5f97\u306a\u5834\u5408\u304c\u3042\u308a\u307e\u3059\u3002<\/p>\n

\u307e\u3068\u3081<\/h2>\n

SYBR Green\u3067\u89e3\u96e2\u66f2\u7dda\u306b\u8907\u6570\u306e\u30d4\u30fc\u30af\u304c\u307f\u3089\u308c\u308b\u5834\u5408\u306f\u3001\u975e\u7279\u7570\u7684\u306a\u5897\u5e45\u304c\u8d77\u304d\u3066\u304a\u308a\u3001\u5b9a\u91cf\u7684\u306a\u7d50\u679c\u304c\u5f97\u3089\u308c\u3066\u3044\u307e\u305b\u3093\u3002\u307e\u305f\u3001\u30d7\u30e9\u30a4\u30de\u30fc\u6fc3\u5ea6\u3084Annealing\uff0fExtension\u306e\u6e29\u5ea6\u6761\u4ef6\u306e\u6700\u9069\u5316\u306b\u3088\u3063\u3066\u6539\u5584\u3059\u308b\u5834\u5408\u3082\u3042\u308a\u307e\u3059\u3002\u305d\u308c\u3067\u3082\u30c0\u30e1\u306a\u5834\u5408\u306f\u601d\u3044\u5207\u3063\u3066\u30d7\u30e9\u30a4\u30de\u30fc\u3092\u518d\u8a2d\u8a08\u3059\u308b\u3053\u3068\u3092\u30aa\u30b9\u30b9\u30e1\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

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

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

 <\/p>\n

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