{"id":9964,"date":"2021-09-10T20:27:27","date_gmt":"2021-09-10T11:27:27","guid":{"rendered":"https:\/\/admin.acceleratingscience.com\/learning-at-the-bench\/?p=9964"},"modified":"2026-02-10T14:57:28","modified_gmt":"2026-02-10T05:57:28","slug":"rox_workshop_gsd_ts_1","status":"publish","type":"post","link":"https:\/\/www.thermofisher.com\/blog\/learning-at-the-bench\/rox_workshop_gsd_ts_1\/","title":{"rendered":"\u3010\u3084\u3063\u3066\u307f\u305f\u3011\u30ea\u30a2\u30eb\u30bf\u30a4\u30e0PCR\u306e\u53cd\u5fdc\u6db2\u5206\u6ce8\u30d4\u30da\u30c3\u30c6\u30a3\u30f3\u30b0\u8aa4\u5dee\u3092ROX\u88dc\u6b63\u3057\u3066\u307f\u305f"},"content":{"rendered":"

2021\u5e741\u6708\uff5e3\u6708\u306b\u958b\u50ac\u3057\u305f\u300c\u3084\u3063\u3066\u307f\u305f\u30b7\u30ea\u30fc\u30ba\u3000\u30a2\u30a4\u30c7\u30a2\u52df\u96c6\u30a4\u30d9\u30f3\u30c8\u300d\u306b\u591a\u304f\u306e\u3054\u5fdc\u52df\u3092\u3044\u305f\u3060\u304d\u3042\u308a\u304c\u3068\u3046\u3054\u3056\u3044\u307e\u3057\u305f\u3002\u7686\u3055\u307e\u304b\u3089\u9802\u3044\u305f\u30a2\u30a4\u30c7\u30a2\u306b\u3064\u3044\u3066\u3001\u793e\u5185\u3067\u691c\u8a0e\u3057\u3001Blog\u306e\u8a18\u4e8b\u3068\u3057\u3066\u516c\u958b\u3059\u308b\u30c6\u30fc\u30de\u3092\u6c7a\u5b9a\u3057\u307e\u3057\u305f\uff01\uff08\u5f53\u9078\u767a\u8868\u306e\u8a18\u4e8b\u306f\u3053\u3061\u3089<\/a>\uff09
\n\u4eca\u56de\u306f2\u3064\u76ee\u306eROX\u88dc\u6b63\u306b\u3064\u3044\u3066\u300c\u3084\u3063\u3066\u307f\u305f\u5b9f\u9a13\u300d\u304c\u5b8c\u4e86\u3057\u305f\u306e\u3067\u3001\u305d\u306e\u7d50\u679c\u3092\u516c\u958b\u3057\u307e\u3059\u3002<\/p>\n

1\u3064\u76ee\u306e\u30c8\u30e9\u30f3\u30b9\u30d5\u30a7\u30af\u30b7\u30e7\u30f3\u8a66\u85ac\u306e\u30a4\u30f3\u30ad\u30e5\u30d9\u30fc\u30b7\u30e7\u30f3\u6642\u9593\u306b\u3064\u3044\u3066\u306e\u8a18\u4e8b\u306f\u3053\u3061\u3089<\/a>\u3092\u3054\u89a7\u304f\u3060\u3055\u3044\u3002<\/p>\n

\u306f\u3058\u3081\u306b<\/h2>\n

\u3053\u306e\u4f01\u753b\u3067\u4eca\u56de\u304a\u5fdc\u3048\u3059\u308b\u3053\u3068\u306b\u306a\u3063\u305f\u306e\u306f\u3001\u30cf\u30f3\u30c9\u30eb\u30cd\u30fc\u30e0 “Narupiyo” \u3055\u3093\u304b\u3089\u3054\u5fdc\u52df\u3044\u305f\u3060\u3044\u305f\u30a2\u30a4\u30c7\u30a2\u3067\u3059\u3002<\/p>\n

\u300c\u30ea\u30a2\u30eb\u30bf\u30a4\u30e0PCR ROX\u306f\u3069\u3053\u307e\u3067\u8aa4\u5dee\u3092\u88dc\u6b63\u3057\u3066\u304f\u308c\u308b\u306e\u304b\u691c\u8a3c\u3057\u3066\u307f\u305f\u300d \u3069\u306e\u7a0b\u5ea6\u307e\u3067\u30d4\u30da\u30c3\u30c6\u30a3\u30f3\u30b0\u8aa4\u5dee\u3092\u88dc\u6b63\u3057\u3066\u304f\u308c\u3066\u3044\u308b\u306e\u3067\u3057\u3087\u3046\u304b\uff0e\uff08\u539f\u6587\u307e\u307e\uff09\u3002<\/p>\n

\u3053\u308c\u3082\u826f\u3044\u30a2\u30a4\u30c7\u30a2\u3067\u3059\u306d\uff01ROX\u88dc\u6b63\u6a5f\u80fd\u306f\u5f0a\u793e\u306e\u30ea\u30a2\u30eb\u30bf\u30a4\u30e0PCR\u30b7\u30b9\u30c6\u30e0\u306e\u7279\u9577\u3067\u3059\u306e\u3067\u3001\u308f\u308c\u308f\u308c\u304c\u3084\u3089\u306d\u3070\u8ab0\u304c\u3084\u308b\uff01\u3068\u3044\u3046\u610f\u6c17\u8fbc\u307f\u3067\u5b9f\u969b\u306b\u5b9f\u9a13\u3057\u3066\u307f\u308b\u3053\u3068\u306b\u3057\u307e\u3057\u305f\u3002ROX\u88dc\u6b63\u3068\u306f\u3001\u53cd\u5fdc\u6db2\u4e2d\u306b\u542b\u307e\u308c\u3066\u3044\u308b\u30ea\u30d5\u30a1\u30ec\u30f3\u30b9\u3068\u306a\u308bROX\u8272\u7d20\u306e\u86cd\u5149\u5f37\u5ea6\u3067\u3001\u30ec\u30dd\u30fc\u30bf\u30fc\u8272\u7d20\uff08e.g. FAM, VIC, or SYBR\u2122 Green\uff09\u306e\u86cd\u5149\u5f37\u5ea6\u3092\u5272\u308b\u3053\u3068\u306b\u3088\u308a\u30a6\u30a7\u30eb\u9593\u306e\u3070\u3089\u3064\u304d\u3092\u6a19\u6e96\u5316\u3059\u308b\u3053\u3068\u3067\u3059\u3002ROX\u88dc\u6b63\u306b\u3064\u3044\u3066\u52d5\u753b<\/a>\u3082\u3054\u7528\u610f\u3057\u3066\u304a\u308a\u307e\u3059\u306e\u3067\u3001\u305c\u3072\u3054\u89a7\u304f\u3060\u3055\u3044\u3002<\/p>\n

\u6750\u6599\u3068\u65b9\u6cd5<\/h2>\n

\u6750\u6599\uff1a<\/p>\n\n\n\n\n\n\n\n\n
\u30ea\u30a2\u30eb\u30bf\u30a4\u30e0PCR\u53cd\u5fdc\u6db2<\/strong><\/td>\n(\u03bcL)<\/td>\n<\/tr>\n
HeLa cDNA\uff081 ng\/\u00b5L\uff09<\/td>\n80<\/td>\n<\/tr>\n
Applied Biosystems\u2122 TaqMan\u2122 Fast Advanced Master Mix<\/td>\n400<\/td>\n<\/tr>\n
Applied Biosystems\u2122 TaqMan Gene Expression Assay (20x) GAPDH<\/td>\n40<\/td>\n<\/tr>\n
Nuclease-Free Water<\/td>\n280<\/td>\n<\/tr>\n
Total<\/td>\n800<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

\u203b\u30a6\u30a7\u30eb \u9593\u3067\u8abf\u88fd\u8aa4\u5dee\u304c\u751f\u3058\u306a\u3044\u3088\u3046\u306b\u3059\u308b\u305f\u3081\u3001\u5c11\u3057\u591a\u3081\u306e40 well \u5206\u3092\u307e\u3068\u3081\u3066\u8abf\u88fd\u3057\u3001\u5404\u5bb9\u91cf\u3092\u5206\u6ce8\u3057\u307e\u3057\u305f\u3002<\/p>\n

\u65b9\u6cd5\uff1a
\n1 \u30a6\u30a7\u30eb \u3042\u305f\u308a\u306e\u6a19\u6e96\u306e\u53cd\u5fdc\u6db2\u91cf\u306f20 \u00b5L\u306b\u8a2d\u5b9a\u3057\u307e\u3057\u305f\u3002\u30d4\u30da\u30c3\u30c6\u30a3\u30f3\u30b0\u8aa4\u5dee\u306b\u3088\u308b\u5f71\u97ff\u3092\u898b\u308b\u305f\u3081\u3001\u3042\u3048\u3066\u6db2\u91cf\u3092\u00b110 \u00b5L\u3068\u632f\u3063\u3066\u300110 \u00b5L\/\u30a6\u30a7\u30eb\u306830 \u00b5L\/\u30a6\u30a7\u30eb\u306e\u3082\u306e\u3068\u6bd4\u8f03\u3057\u307e\u3057\u305f\uff08\u56f31\uff09\u3002\u3082\u3061\u308d\u3093\u3001\u5404\u8a66\u85ac\u306e\u6bd4\u7387\u306f\u5171\u901a\u306b\u3057\u3066\u3001Total \u306e\u6db2\u91cf\u306e\u307f\u3092\u5909\u5316\u3055\u305b\u307e\u3057\u305f\u3002\u30ea\u30a2\u30eb\u30bf\u30a4\u30e0PCR\u30b7\u30b9\u30c6\u30e0\u306f\u3001Applied Biosystems\u2122 QuantStudio\u2122 5<\/a> \u3092\u4f7f\u7528\u3057\u3001Fast mode\u306b\u8a2d\u5b9a\u3057\u3066\u4e0b\u8a18\u6761\u4ef6\u3067\u30e9\u30f3\u3092\u5b9f\u65bd\u3057\u307e\u3057\u305f\u3002
\n\"\"
\nROX\u88dc\u6b63\u306e\u6709\u7121\u306b\u304a\u3044\u3066\u3001\u5897\u5e45\u66f2\u7dda\u3068CT<\/sub>\u5024\u3092\u6bd4\u8f03\u3057\u307e\u3057\u305f\uff08n = 12\uff09\u3002
\n\"\"<\/p>\n

\u7d50\u679c\u3068\u8003\u5bdf<\/h2>\n

\u305d\u308c\u3067\u306f\u30d4\u30da\u30c3\u30c6\u30a3\u30f3\u30b0\u8aa4\u5dee\u3092ROX\u88dc\u6b63\u3057\u3066\u307f\u305f\u7d50\u679c\u3092\u3054\u7d39\u4ecb\u3057\u307e\u3059\u3002<\/p>\n

\u307e\u305a\u3001ROX\u88dc\u6b63\u3092\u3057\u306a\u3044\u5834\u5408\u306e\u7d50\u679c\u3092\u78ba\u8a8d\u3057\u307e\u3057\u3087\u3046\u3002\u53cd\u5fdc\u6db2\u91cf\u304c10 \u00b5L(\u8d64\u7dda)\u300120 \u00b5L\uff08\u9752\u7dda\uff09\u300130 \u00b5L\uff08\u7dd1\u7dda\uff09 \u3067\u306f\u3063\u304d\u308a\u7570\u306a\u308b\u5897\u5e45\u66f2\u7dda\u306b\u306a\u308a\u307e\u3057\u305f\uff08\u56f32\u5de6\uff09\u3002\u53cd\u5fdc\u6db2\u91cf\u304c\u591a\u3044\u307b\u3069CT<\/sub>\u5024\u203b<\/sup>1<\/sup> \u304c\u4f4e\u304f\u306a\u3063\u305f\u3053\u3068\u304b\u3089\uff08\u88681\uff09\u3001\u30d4\u30da\u30c3\u30c6\u30a3\u30f3\u30b0\u8aa4\u5dee\u3067\u53cd\u5fdc\u6db2\u91cf\u3092\u591a\u304f\u52a0\u3048\u3066\u3057\u307e\u3046\u3068\u3001\u907a\u4f1d\u5b50\u767a\u73fe\u91cf\u306a\u3069\u3092\u771f\u306e\u5024\u3088\u308a\u3082\u591a\u3081\u306b\u5b9a\u91cf\u3057\u3066\u3057\u307e\u3046\u3053\u3068\u304c\u8003\u3048\u3089\u308c\u307e\u3057\u305f\u3002\u9006\u306b\u3001\u53cd\u5fdc\u6db2\u91cf\u304c\u5c11\u306a\u3044\u5834\u5408\u306fCT<\/sub>\u5024\u304c\u9ad8\u304f\u306a\u3063\u305f\u306e\u3067\u3001\u907a\u4f1d\u5b50\u767a\u73fe\u91cf\u306a\u3069\u3092\u5c11\u306a\u3081\u306b\u5b9a\u91cf\u3057\u3066\u3057\u307e\u3046\u30a8\u30e9\u30fc\u304c\u751f\u3058\u308b\u53ef\u80fd\u6027\u304c\u793a\u5506\u3055\u308c\u307e\u3059\u3002\u3055\u3089\u306b\u3001\u4eca\u56de\u306f12\u53cd\u5fa9\u3067\u5b9f\u9a13\u3092\u5b9f\u65bd\u3057\u307e\u3057\u305f\u304c\u3001\u8d64\u30fb\u9752\u30fb\u7dd1\u306e\u305d\u308c\u305e\u308c\u306e\u7dda\u304c\u3074\u3063\u305f\u308a\u91cd\u306a\u3089\u306a\u3044\u3053\u3068\u304b\u3089\u3001\u53cd\u5fdc\u6db2\u91cf\u306e\u9055\u3044\u306e\u307f\u306a\u3089\u305a \u30a6\u30a7\u30eb \u3054\u3068\u306b\u8aa4\u5dee\u304c\u3042\u308b\u3053\u3068\u3082\u660e\u78ba\u306b\u8a8d\u8b58\u3067\u304d\u307e\u3057\u305f\uff08\u56f32\u5de6\uff09\u3002<\/p>\n

\u6b21\u306b\u3001ROX\u88dc\u6b63\u3057\u305f\u5834\u5408\u306e\u7d50\u679c\u3092\u78ba\u8a8d\u3057\u307e\u3057\u3087\u3046\u3002\u3071\u3063\u3068\u898b\u308b\u3068\u8d64\u3044\u7dda\u304c1\u672c\u3060\u3051\u306e\u3088\u3046\u306b\u601d\u3048\u307e\u3059\uff08\u56f32\u53f3\uff09\u3002\u5b9f\u306f\u3001\u3053\u308c\u304cROX\u88dc\u6b63\u306e\u52b9\u679c\u3092\u3088\u304f\u73fe\u3057\u3066\u3044\u307e\u3059\u3002\u3053\u306e\u56f3\u306f1 \u30a6\u30a7\u30eb \u3060\u3051\u306e\u7d50\u679c\u3092\u8868\u793a\u3057\u3066\u3044\u308b\u308f\u3051\u3067\u306f\u306a\u304f\u300110 \u00b5L\u300120 \u00b5L\u300130 \u00b5L\u305d\u308c\u305e\u308c12\u53cd\u5fa9\u306736\u672c\u306e\u5897\u5e45\u66f2\u7dda\u3092\u8868\u793a\u3057\u3066\u3044\u307e\u3059\u3002\u3057\u304b\u3057\u3001\u307b\u307c\u5b8c\u5168\u306b\u91cd\u306a\u3063\u3066\u3044\u308b\u305f\u3081\u30011\u672c\u306e\u8d64\u3044\u7dda\u306e\u3088\u3046\u306b\u3057\u304b\u898b\u3048\u306a\u3044\u3068\u3044\u3046\u3053\u3068\u3067\u3059\u3002\u3064\u307e\u308a\u3001\u53cd\u5fdc\u6db2\u91cf\u5dee\u3084\u30a6\u30a7\u30eb \u3054\u3068\u306e\u5dee\u304c\u539f\u56e0\u3068\u306a\u3063\u306636\u672c\u306b\u5206\u304b\u308c\u3066\u898b\u3048\u3066\u3044\u305f\u5897\u5e45\u66f2\u7dda\u3067\u3059\u304c\uff08\u56f32\u5de6\uff09\u3001\u30c7\u30fc\u30bf\u3092ROX\u88dc\u6b63\u3059\u308b\u3053\u3068\u3067\u8aa4\u5dee\u304c\u5c0f\u3055\u304f\u306a\u3063\u30661\u672c\u306b\u91cd\u306a\u308a\u3001\u3059\u3079\u3066\u306e\u30a6\u30a7\u30eb\u3067\u3070\u3089\u3064\u304d\u306e\u5c0f\u3055\u3044CT<\/sub>\u5024\u3092\u5f97\u308b\u3053\u3068\u304c\u3067\u304d\u307e\u3057\u305f\uff08\u88681\uff09\u3002<\/p>\n

ROX\u306e\u6709\u7121\u3092\u5177\u4f53\u7684\u306b\u6bd4\u8f03\u3059\u308b\u3068\u3001\u53cd\u5fdc\u6db2\u91cf\u306e\u5dee\u304c\u3042\u3063\u3066\u3082CT<\/sub>\u5024\u306e\u5dee\u304c\u5c0f\u3055\u304f\u306a\u308a\u300112\u53cd\u5fa9\u5185\u306e\u3070\u3089\u3064\u304d\uff08\u6a19\u6e96\u504f\u5dee\uff09\u3082\u5c0f\u3055\u304f\u306a\u308a\u307e\u3057\u305f\uff08\u88681\uff09\u3002\u3053\u306e\u3053\u3068\u306f\u3001\u305f\u3068\u3048\u30d4\u30da\u30c3\u30c6\u30a3\u30f3\u30b0\u8aa4\u5dee\u304c\u5927\u304d\u304f\u3066\u3082\u3001ROX\u88dc\u6b63\u306b\u3088\u308a\u3070\u3089\u3064\u304d\u306e\u5c11\u306a\u3044\u5b89\u5b9a\u3057\u305f\u5b9a\u91cf\u5024\u304c\u5f97\u3089\u308c\u308b\u3053\u3068\u3092\u793a\u3057\u3066\u3044\u307e\u3059\u3002\u4eca\u56de\u3054\u5fdc\u52df\u3044\u305f\u3060\u3044\u305f \u201cNarupiyo\u201d \u3055\u3093\u306b\u306f\u3001\u300c\u00b110 \u00b5L\u307e\u3067\u306e\u30d4\u30da\u30c3\u30c6\u30a3\u30f3\u30b0\u8aa4\u5dee\u3067\u3042\u308c\u3070\u3001\u304b\u306a\u308a\u7cbe\u5ea6\u9ad8\u304fROX\u88dc\u6b63\u3059\u308b\u3053\u3068\u304c\u53ef\u80fd\u3067\u3059\u300d\u3068\u304a\u7b54\u3048\u3059\u308b\u3053\u3068\u304c\u3067\u304d\u305d\u3046\u3067\u3059\u3002<\/p>\n

\u203b1 CT<\/sub>\u5024\uff1aThreshold Cycle \u306e\u6570\u5024\u306e\u3053\u3068\u3067\u3001Threshold Line\u3068\u5897\u5e45\u66f2\u7dda\u306e\u4ea4\u70b9\u306e\u30b5\u30a4\u30af\u30eb\u6570\u306e\u3053\u3068\u3002<\/p>\n

\"\"<\/p>\n

\"\"<\/p>\n

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