{"id":12689,"date":"2022-10-25T09:10:24","date_gmt":"2022-10-25T00:10:24","guid":{"rendered":"https:\/\/admin.acceleratingscience.com\/learning-at-the-bench\/?p=12689"},"modified":"2025-03-10T12:43:32","modified_gmt":"2025-03-10T03:43:32","slug":"rna_workshop_bid_ts_3","status":"publish","type":"post","link":"https:\/\/www.thermofisher.com\/blog\/learning-at-the-bench\/rna_workshop_bid_ts_3\/","title":{"rendered":"\u3010\u3084\u3063\u3066\u307f\u305f\u3011RNA\u62bd\u51fa\u306b\u30af\u30ed\u30ed\u30db\u30eb\u30e0\u3092\u4f7f\u308f\u305a\u306b\u30d6\u30ed\u30e2\u30af\u30ed\u30ed\u30d7\u30ed\u30d1\u30f3\u3067\u4ee3\u66ff\u3057\u3066\u307f\u305f"},"content":{"rendered":"

RNA\u62bd\u51fa\u65b9\u6cd5\u306e1\u3064\u306b\u30d5\u30a7\u30ce\u30fc\u30eb\u30fb\u30af\u30ed\u30ed\u30db\u30eb\u30e0\u62bd\u51fa\u304c\u3042\u308a\u307e\u3059\u3002\u30d5\u30a7\u30ce\u30fc\u30eb\u3082\u30af\u30ed\u30ed\u30db\u30eb\u30e0\u3082\u3069\u3061\u3089\u3082\u5287\u7269\u306b\u6307\u5b9a\u3055\u308c\u3066\u304a\u308a\u3001\u4f7f\u7528\u8005\u306e\u5b89\u5168\u6027\u306e\u9762\u304b\u3089\u3082\u306a\u308b\u3079\u304f\u4f7f\u7528\u3092\u63a7\u3048\u305f\u3044\u8a66\u85ac\u3067\u306f\u306a\u3044\u3067\u3057\u3087\u3046\u304b\u3002<\/p>\n

\u4eca\u56de\u306f\u30af\u30ed\u30ed\u30db\u30eb\u30e0\u306e\u4ee3\u308f\u308a\u306b1-\u30d6\u30ed\u30e2-3-\u30af\u30ed\u30ed\u30d7\u30ed\u30d1\u30f3\u304c\u4f7f\u7528\u53ef\u80fd\u306a\u3053\u3068\u3092\u3054\u7d39\u4ecb\u3057\u307e\u3059\u3002<\/p>\n

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

\u30af\u30ed\u30ed\u30db\u30eb\u30e0\u306f\u4e2d\u67a2\u795e\u7d4c\u306b\u4f5c\u7528\u3059\u308b\u533b\u85ac\u7528\u5916\u5287\u7269\u3067\u3001\u5b9f\u9a13\u5ba4\u3067\u306f\u30c9\u30e9\u30d5\u30c8\u5185\u3067\u4f7f\u7528\u3059\u308b\u3053\u3068\u304c\u5f37\u304f\u5968\u52b1\u3055\u308c\u3001\u6cd5\u6539\u6b63\u306b\u3088\u308a\u53d6\u308a\u6271\u3044\u7ba1\u7406\u304c\u3088\u308a\u53b3\u3057\u304f\u306a\u3063\u3066\u3044\u307e\u3059\u3002\u4e3b\u8981\u306aRNA\u62bd\u51fa\u65b9\u6cd5\u306e1\u3064\u3067\u3042\u308bAGPC\uff08Acid Guanidinium Thiocyanate-Phenol-Chloroform extraction\uff09\u6cd5\u3067\u306f\u3001\u30d5\u30a7\u30ce\u30fc\u30eb\u30fb\u30af\u30ed\u30ed\u30db\u30eb\u30e0\u62bd\u51fa\u306b\u3088\u308a\u3001\u30b2\u30ce\u30e0DNA\u3084\u30bf\u30f3\u30d1\u30af\u8cea\u306a\u3069\u306e\u593e\u96d1\u6210\u5206\u3092\u52b9\u679c\u7684\u306b\u9664\u53bb\u3057\u307e\u3059\u3002\u4ee5\u524d\u3088\u308a\u30af\u30ed\u30ed\u30db\u30eb\u30e0\u306e\u4ee3\u66ff\u3048\u8a66\u85ac\u3068\u3057\u30661-\u30d6\u30ed\u30e2-3-\u30af\u30ed\u30ed\u30d7\u30ed\u30d1\u30f3\uff08CAS RN:109-70-6\u3001\u4ee5\u964d\u3001BCP\uff09\u304c\u7d39\u4ecb\u3055\u308c\u3066\u304a\u308a\u3001\u30af\u30ed\u30ed\u30db\u30eb\u30e0\u3088\u308a\u4f4e\u6bd2\u6027\u3067\u3001\u3055\u3089\u306b\u6dfb\u52a0\u91cf\u304c\u534a\u91cf\u3067\u826f\u3044\u305f\u3081\u64cd\u4f5c\u6027\u306b\u3082\u305f\u3051\u3066\u3044\u308b\u3068\u7d39\u4ecb\u3055\u308c\u3066\u3044\u307e\u3059\u3002\u672c\u7a3f\u3067\u306fBCP\u306e\u6709\u52b9\u6027\u306b\u3064\u3044\u3066\u691c\u8a3c\u5b9f\u9a13\u3092\u884c\u3044\u3001\u305d\u306e\u6709\u52b9\u6027\u306b\u3064\u3044\u3066\u78ba\u8a8d\u3092\u884c\u3044\u307e\u3057\u305f\u3002<\/p>\n

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

RNA\u62bd\u51fa\u306e\u30b5\u30f3\u30d7\u30eb\u306f\u3001\u30de\u30a6\u30b9\u306e\u809d\u81d3\u3092\u7d30\u5207\u308c\u306b\u3057\u3066Invitrogen\u2122 RNAlater\u2122 Stabilization Solution<\/a>\u306b\u6d78\u6f2c\u3057-20\u2103\u30675\u5e74\u4ee5\u4e0a\u4fdd\u5b58\u3057\u3066\u3044\u305f\u3082\u306e\u3092\u7528\u3044\u307e\u3057\u305f\u3002\u30de\u30a6\u30b9\u809d\u81d320 mg\u306b\u3001AGPC\u6cd5\u3092\u30d9\u30fc\u30b9\u3068\u3057\u305fRNA\u62bd\u51fa\u8a66\u85ac\u3067\u3042\u308bInvitrogen\u2122 TRIzol\u2122 Reagent<\/a>\u3092200 \u00b5L\u52a0\u3048\u3066\u3001Fisherbrand\u2122 RNase-Free Disposable Pellet Pestles<\/a> \u3092\u7528\u3044\u3066\u3088\u304f\u3059\u308a\u3064\u3076\u3057\u307e\u3057\u305f\u3002\u30b5\u30f3\u30d7\u30eb\u30a4\u30f3\u30d7\u30c3\u30c8\u91cf\u306b\u8d77\u56e0\u3059\u308b\u8aa4\u5dee\u3092\u9664\u304f\u305f\u3081\u3001\u30db\u30e2\u30b8\u30ca\u30a4\u30ba\u3057\u305f\u3082\u306e\u3092\u30af\u30ed\u30ed\u30db\u30eb\u30e0\u7528\u3068BCP\u7528\u306e2\u3064\u306b\u7b49\u91cf\u305a\u3064\u5206\u3051\u3066\u3001RNA\u62bd\u51fa\u64cd\u4f5c\u3092\u7d99\u7d9a\u3057\u307e\u3057\u305f\uff08n=3\uff09\u3002\u5404\u30c1\u30e5\u30fc\u30d6\u306b900 \u00b5L\u306eTRIzol\u6eb6\u6db2\u3092\u52a0\u3048\u3066\uff08Total 1000 mL\uff09\u30db\u30e2\u30b8\u30ca\u30a4\u30ba\u3057\u307e\u3057\u305f\u3002\u5ba4\u6e29\u30675 min\u304a\u3044\u3066\u6838\u9178\u7d50\u5408\u30bf\u30f3\u30d1\u30af\u8cea\u306a\u3069\u3092\u5909\u6027\u3055\u305b\u307e\u3057\u305f\u3002<\/p>\n

BCP\u306f\u30af\u30ed\u30ed\u30db\u30eb\u30e0\u306e\u534a\u91cf\u3092\u4f7f\u7528\u3057\u307e\u3059\u306e\u3067\u3001\u30af\u30ed\u30ed\u30db\u30eb\u30e0\u4f7f\u7528\u306e\u30b5\u30f3\u30d7\u30eb\u306b\u306f200 \u00b5L\u3001BCP\u4f7f\u7528\u306e\u30b5\u30f3\u30d7\u30eb\u306b\u306f100 \u00b5L\u52a0\u3048\u3001\u305d\u308c\u305e\u308cTotal 1200 \u00b5L\u3082\u3057\u304f\u306fTotal 1100 \u00b5L\u306b\u3057\u307e\u3057\u305f\u300215 sec\u624b\u3067\u6fc0\u3057\u304f\u632f\u308b\u3053\u3068\u3067\u6df7\u5408\u3057\u3001\u5ba4\u6e29\u30672 min\u9759\u7f6e\u3057\u307e\u3057\u305f\u300212,000 g\u300115 min\u30014\u2103\u3067\u9060\u5fc3\u5f8c\u3001\u6c34\u76f8400 \u00b5L\u3092\u65b0\u3057\u3044\u30c1\u30e5\u30fc\u30d6\u306b\u56de\u53ce\u3057\u307e\u3057\u305f\u3002\u5404\u30c1\u30e5\u30fc\u30d6\u306b500 \u00b5L\u306e\u30a4\u30bd\u30d7\u30ed\u30d1\u30ce\u30fc\u30eb\u3092\u52a0\u3048\u5ba4\u6e29\u306710 min\u9759\u7f6e\u3057\u305f\u5f8c\u300112,000 g\u300110 min\u30014\u2103\u3067\u9060\u5fc3\u3057\u3066RNA\u3092\u56de\u53ce\u3057\u307e\u3057\u305f\u3002
\n\u56de\u53ce\u3057\u305fRNA\u306f70%\u30a8\u30bf\u30ce\u30fc\u30eb\u3067\u4e00\u5ea6\u30ea\u30f3\u30b9\u3057\u30017,500 g\u30015 min\u30014\u2103\u3067\u6c88\u6bbf\u3055\u305b\u305f\u5f8c\u306b\u98a8\u4e7e\u3055\u305b\u3001\u6700\u7d42\u7684\u306b\u306f50 \u00b5L\u306eInvitrogen\u2122 Nuclease-Free Water<\/a> \u3067\u6eb6\u89e3\u3057\u307e\u3057\u305f\u3002\u62bd\u51fa\u3057\u305fRNA\u306e\u8a55\u4fa1\u306f\u3001Thermo Scientific\u2122 NanoDrop\u2122 OneC<\/a>\u304a\u3088\u3073Agilent\u793eBioanalyzer\uff08Agilent RNA6000\u30d4\u30b3\u30ad\u30c3\u30c8\uff09\u3092\u7528\u3044\u3066\u6e2c\u5b9a\u3059\u308b\u3053\u3068\u3067\u5b9f\u65bd\u3057\u307e\u3057\u305f\u3002<\/p>\n

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

\u4e0a\u8a18\u306e\u65b9\u6cd5\u3067\u3001\u30af\u30ed\u30ed\u30db\u30eb\u30e0\u3068BCP\u3092\u6bd4\u8f03\u3057\u307e\u3057\u305f\u3002\u7d50\u679c\u306f\u3001\u3069\u3061\u3089\u306e\u8a66\u85ac\u3092\u4f7f\u7528\u3057\u3066\u3082\u3001\u554f\u984c\u306a\u304fRNA\u3092\u62bd\u51fa\u3059\u308b\u3053\u3068\u304c\u3067\u304d\u307e\u3057\u305f\uff08\u88681\uff09\u3002\u30af\u30ed\u30ed\u30db\u30eb\u30e0\u3092BCP\u306b\u7f6e\u304d\u63db\u3048\u308b\u3053\u3068\u306b\u3088\u308a\u3001\u4e2d\u9593\u5c64\u306e\u593e\u96d1\u6210\u5206\u304c\u5bc6\u306b\u51dd\u96c6\u3055\u308c\u307e\u3059\u3002\u3053\u308c\u306fBCP\u304c\u30af\u30ed\u30ed\u30db\u30eb\u30e0\u3088\u308a\u3082\u758e\u6c34\u6027\u306e\u9ad8\u3044\u6709\u6a5f\u6eb6\u5a92\u3067\u3042\u308b\u305f\u3081\u3068\u8003\u3048\u3089\u308c\u307e\u3059\u3002\u3053\u308c\u306b\u3088\u308a\u4e0a\u6e05\u306e\u56de\u53ce\u64cd\u4f5c\u304c\u3057\u3084\u3059\u304f\u306a\u308a\u3001\u4e0d\u6eb6\u6027\u306e\u593e\u96d1\u6210\u5206\u306e\u30b3\u30f3\u30bf\u30df\u30cd\u30fc\u30b7\u30e7\u30f3\u3092\u9632\u3050\u3053\u3068\u304c\u3067\u304d\u308b\u306e\u3067\u3001\u30a4\u30bd\u30d7\u30ed\u30d1\u30ce\u30fc\u30eb\u6c88\u6bbf\u5f8c\u306eRNA\u30da\u30ec\u30c3\u30c8\u306e\u6eb6\u89e3\u304c\u30b9\u30e0\u30fc\u30ba\u306b\u306a\u308b\u3053\u3068\u304c\u4e88\u60f3\u3055\u308c\u307e\u3059\u3002\u3053\u306e\u305f\u3081\u306bRNA\u306e\u53ce\u91cf\u304c\u82e5\u5e72\u9ad8\u304b\u3063\u305f\u306e\u3060\u3068\u8003\u3048\u3089\u308c\u307e\u3057\u305f\u3002<\/p>\n

\u3055\u3089\u306b\u3001\u6fc3\u5ea6\u4ee5\u5916\u306eRNA\u54c1\u8cea\u306e\u6307\u6a19\u3068\u3057\u3066\u3001A260\/A280\u3001A260\/A230\u3001RIN\u3092\u6bd4\u8f03\u3057\u307e\u3057\u305f\u304c\u3001\u4f7f\u7528\u3057\u305f\u8a66\u85ac\u306e\u9055\u3044\u3067\u5927\u304d\u306a\u5dee\u306f\u898b\u3089\u308c\u307e\u305b\u3093\u3067\u3057\u305f\uff08\u88681\uff09\u3002\u3053\u306e\u7d50\u679c\u304b\u3089\u3001TRIzol\u2122 Reagent<\/a>\u3092\u4f7f\u3063\u3066RNA\u62bd\u51fa\u3059\u308b\u969b\u3001\u5f93\u6765\u306f\u30af\u30ed\u30ed\u30db\u30eb\u30e0\u3092\u4f7f\u7528\u3059\u308b\u30d7\u30ed\u30c8\u30b3\u30eb\u3067\u3059\u304c\u3001\u534a\u91cf\u306eBCP\u3067\u3082\u4ee3\u66ff\u53ef\u80fd\u3067\u3042\u308b\u3053\u3068\u3092\u304a\u793a\u3057\u3067\u304d\u307e\u3057\u305f\u3002<\/p>\n

\"RNA\u62bd\u51fa\"<\/p>\n

\u3044\u304b\u304c\u3067\u3057\u305f\u3067\u3057\u3087\u3046\u304b\u3002<\/p>\n

\u4eca\u56de\u306f\u3001TRIzol Reagent\u3092\u4f7f\u3063\u3066RNA\u62bd\u51fa\u3059\u308b\u969b\u306b\u3001\u30af\u30ed\u30ed\u30db\u30eb\u30e0\u3092BCP\u3067\u4ee3\u66ff\u53ef\u80fd\u3067\u3042\u308b\u3053\u3068\u3092\u3054\u7d39\u4ecb\u3057\u307e\u3057\u305f\u3002\u5287\u7269\u3067\u3042\u308b\u30af\u30ed\u30ed\u30db\u30eb\u30e0\u306e\u4f7f\u7528\u91cf\u306e\u524a\u6e1b\u3084\u3001\u4f7f\u7528\u8005\u306e\u5b89\u5168\u6027\u306e\u78ba\u4fdd\u306e\u305f\u3081\u306b\u3001BCP\u306e\u4f7f\u7528\u3092\u691c\u8a0e\u3055\u308c\u3066\u3082\u826f\u3044\u304b\u3082\u3057\u308c\u307e\u305b\u3093\u3002<\/p>\n

\u5f53\u793e\u3067\u306fRNA\u62bd\u51fa\u3084\u30ea\u30a2\u30eb\u30bf\u30a4\u30e0PCR\u3001\u4ed6\u306b\u3082\u7d30\u80de\u57f9\u990a\u3001\u30a6\u30a7\u30b9\u30bf\u30f3\u30d6\u30ed\u30c3\u30c6\u30a3\u30f3\u30b0\u306a\u3069\u3001\u5b9f\u969b\u306b\u5b9f\u9a13\uff08\u5b9f\u7fd2\uff09\u3092\u884c\u3044\u3064\u3064\u5b66\u3079\u308b\u5404\u7a2e\u30cf\u30f3\u30ba\u30aa\u30f3\u30c8\u30ec\u30fc\u30cb\u30f3\u30b0<\/a>\u3092\u958b\u50ac\u3057\u3066\u3044\u307e\u3059\u3002\u305d\u306e\u4e2d\u3067\u4eca\u56de\u306e\u3088\u3046\u306a\u5b9f\u9a13\u7d50\u679c\u3082\u3054\u7d39\u4ecb\u3057\u3066\u3044\u307e\u3059\u306e\u3067\u3001\u3053\u308c\u304b\u3089\u65b0\u3057\u3044\u5b9f\u9a13\u3092\u59cb\u3081\u3089\u308c\u308b\u65b9\u3001\u3088\u308a\u7406\u89e3\u3092\u6df1\u3081\u305f\u3044\u65b9\u306f\u305c\u3072\u3054\u53c2\u52a0\u304f\u3060\u3055\u3044\uff01<\/p>\n

\u5206\u5b50\u751f\u7269\u5b66\u6559\u5ba4PDF\u7248 \u7121\u6599\u30c0\u30a6\u30f3\u30ed\u30fc\u30c9<\/h3>\n

\u30c0\u30a6\u30f3\u30ed\u30fc\u30c9\u306f\u3053\u3061\u3089<\/a><\/p>\n

 <\/p>\n

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RNA\u62bd\u51fa\u65b9\u6cd5\u306e1\u3064\u306b\u30d5\u30a7\u30ce\u30fc\u30eb\u30fb\u30af\u30ed\u30ed\u30db\u30eb\u30e0\u62bd\u51fa\u304c\u3042\u308a\u307e\u3059\u3002\u30d5\u30a7\u30ce\u30fc\u30eb\u3082\u30af\u30ed\u30ed\u30db\u30eb\u30e0\u3082\u3069\u3061\u3089\u3082\u5287\u7269\u306b\u6307\u5b9a\u3055\u308c\u3066\u304a\u308a\u3001\u4f7f\u7528\u8005\u306e\u5b89\u5168\u6027\u306e\u9762\u304b\u3089\u3082\u306a\u308b\u3079\u304f\u4f7f\u7528\u3092\u63a7\u3048\u305f\u3044\u8a66\u85ac\u3067\u306f\u306a\u3044\u3067\u3057\u3087\u3046\u304b\u3002 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