{"id":569,"date":"2017-02-02T00:00:00","date_gmt":"2017-02-02T00:00:00","guid":{"rendered":"https:\/\/www.thermofisher.com\/blog\/learning-at-the-bench\/guidelines-and-optimizing-for-plasmid-dna-transfection\/"},"modified":"2025-02-26T17:16:28","modified_gmt":"2025-02-26T08:16:28","slug":"guidelines-and-optimizing-for-plasmid-dna-transfection","status":"publish","type":"post","link":"https:\/\/www.thermofisher.com\/blog\/learning-at-the-bench\/guidelines-and-optimizing-for-plasmid-dna-transfection\/","title":{"rendered":"\u30d7\u30e9\u30b9\u30df\u30c9DNA\u30c8\u30e9\u30f3\u30b9\u30d5\u30a7\u30af\u30b7\u30e7\u30f3\u306e\u305f\u3081\u306e\u30ac\u30a4\u30c9\u30e9\u30a4\u30f3\u3068\u6700\u9069\u5316\u65b9\u6cd5"},"content":{"rendered":"

\u53e4\u5178\u7684\u306a\u30c8\u30e9\u30f3\u30b9\u30d5\u30a7\u30af\u30b7\u30e7\u30f3\u6280\u8853\u306f\u3001\u5f53\u521d\u3001\u30d7\u30e9\u30b9\u30df\u30c9DNA\u3092\u7d30\u80de\u306b\u5c0e\u5165\u3059\u308b\u305f\u3081\u306b\u958b\u767a\u3055\u308c\u307e\u3057\u305f\u304c\u3001\u30d7\u30e9\u30b9\u30df\u30c9DNA\u306f\u30c8\u30e9\u30f3\u30b9\u30d5\u30a7\u30af\u30b7\u30e7\u30f3\u306e\u305f\u3081\u306e\u6700\u3082\u4e00\u822c\u7684\u306a\u30d9\u30af\u30bf\u30fc\u3068\u3057\u3066\u73fe\u5728\u3082\u5b58\u5728\u3057\u3066\u3044\u307e\u3059\u3002\u7d44\u63db\u3048\u907a\u4f1d\u5b50\u304a\u3088\u3073\u8abf\u7bc0\u56e0\u5b50\u304c\u542b\u307e\u308c\u308bDNA\u30d7\u30e9\u30b9\u30df\u30c9\u3092\u7d30\u80de\u306b\u5c0e\u5165\u3059\u308b\u3053\u3068\u306b\u3088\u308a\u3001\u907a\u4f1d\u5b50\u306e\u6a5f\u80fd\u304a\u3088\u3073\u8abf\u7bc0\u3001\u907a\u4f1d\u5b50\u7523\u7269\u306e\u5909\u7570\u89e3\u6790\u304a\u3088\u3073\u751f\u7269\u5b66\u7684\u7279\u5fb4\u3001\u907a\u4f1d\u5b50\u767a\u73fe\u306b\u304a\u3051\u308b\u7d30\u80de\u306e\u30e9\u30a4\u30d5\u30b5\u30a4\u30af\u30eb\u3084\u5065\u5eb7\u3078\u306e\u5f71\u97ff\u306b\u95a2\u3059\u308b\u7814\u7a76\u3084\u3001\u7cbe\u88fd\u3057\u30c0\u30a6\u30f3\u30b9\u30c8\u30ea\u30fc\u30e0\u30a2\u30d7\u30ea\u30b1\u30fc\u30b7\u30e7\u30f3\u306b\u4f7f\u7528\u3059\u308b\u305f\u3081\u306e\u30bf\u30f3\u30d1\u30af\u8cea\u306e\u5927\u91cf\u751f\u7523\u304c\u53ef\u80fd\u3067\u3059\u3002<\/p>\n

\u30d9\u30af\u30bf\u30fc\u30b3\u30f3\u30b9\u30c8\u30e9\u30af\u30c8\u306e\u5f62\u614b\uff08\u76f4\u9396\u72b6\u307e\u305f\u306f\u30b9\u30fc\u30d1\u30fc\u30b3\u30a4\u30eb\u72b6\uff09\u304a\u3088\u3073\u30b5\u30a4\u30ba\u3001\u30d7\u30e9\u30b9\u30df\u30c9DNA\u306e\u54c1\u8cea\u3001\u306a\u3089\u3073\u306b\u30d7\u30ed\u30e2\u30fc\u30bf\u30fc\u306e\u9078\u629e\u306f\u3001\u30d7\u30e9\u30b9\u30df\u30c9DNA\u306e\u30c8\u30e9\u30f3\u30b9\u30d5\u30a7\u30af\u30b7\u30e7\u30f3\u52b9\u7387\u306b\u5f71\u97ff\u3092\u4e0e\u3048\u308b\u4e3b\u8981\u306a\u8981\u7d20\u3067\u3059\u3002\u4eca\u56de\u306f\u3001\u30d7\u30e9\u30b9\u30df\u30c9DNA\u3092\u7528\u3044\u305f\u30c8\u30e9\u30f3\u30b9\u30d5\u30a7\u30af\u30b7\u30e7\u30f3\u306b\u3064\u3044\u3066\u306e\u30ac\u30a4\u30c9\u30e9\u30a4\u30f3\u3068\u30d1\u30e9\u30e1\u30fc\u30bf\u306e\u6700\u9069\u5316\u65b9\u6cd5\u306b\u3064\u3044\u3066<\/strong>\u3054\u7d39\u4ecb\u3057\u307e\u3059\u3002<\/p>\n

\u30d7\u30e9\u30b9\u30df\u30c9DNA\u30c8\u30e9\u30f3\u30b9\u30d5\u30a7\u30af\u30b7\u30e7\u30f3\u306e\u30ac\u30a4\u30c9\u30e9\u30a4\u30f3<\/h2>\n

\u30d9\u30af\u30bf\u30fc\u306b\u95a2\u3059\u308b\u8003\u5bdf<\/h3>\n

\u4e00\u904e\u6027\u30c8\u30e9\u30f3\u30b9\u30d5\u30a7\u30af\u30b7\u30e7\u30f3\u306b\u304a\u3044\u3066\u3001\u30b9\u30fc\u30d1\u30fc\u30b3\u30a4\u30ebDNA\u306f\u76f4\u9396\u72b6DNA\u3088\u308a\u3082\u52b9\u7387\u7684\u306b\u53d6\u308a\u8fbc\u307e\u308c\u307e\u3059\u3002<\/strong>\u3053\u308c\u306f\u6050\u3089\u304f\u3001\u74b0\u72b6DNA\u306f\u30a8\u30ad\u30bd\u30cc\u30af\u30ec\u30a2\u30fc\u30bc\u306e\u5f71\u97ff\u3092\u53d7\u3051\u306b\u304f\u3044\u306e\u306b\u5bfe\u3057\u3001\u76f4\u9396\u72b6DNA\u30d5\u30e9\u30b0\u30e1\u30f3\u30c8\u306f\u3053\u306e\u9175\u7d20\u306b\u3088\u3063\u3066\u5373\u5ea7\u306b\u5206\u89e3\u3055\u308c\u308b\u305f\u3081\u3068\u8003\u3048\u3089\u308c\u307e\u3059\uff08Lenachan et al., 2007; von Groll et al., 2006\uff09\u3002\u307e\u305f\u3001\u539f\u5b50\u9593\u529b\u9855\u5fae\u93e1\u89e3\u6790\u3067\u306f\u3001\u74b0\u72b6DNA\u5f62\u614b\u3068\u76f4\u9396\u72b6DNA\u5f62\u614b\u3067\u30ab\u30c1\u30aa\u30f3\u6027\u8102\u8cea\u8a66\u85ac\u3068\u306e\u8907\u5408\u4f53\u5f62\u6210\u30d1\u30bf\u30fc\u30f3\u304c\u975e\u5e38\u306b\u7570\u306a\u308b\u3053\u3068\u304c\u793a\u3055\u308c\u3066\u3044\u307e\u3059\u3002\u3064\u307e\u308a\u3001\u74b0\u72b6DNA\u3067\u306f\u30b3\u30f3\u30d1\u30af\u30c8\u306a\u7403\u72b6\u307e\u305f\u306f\u5186\u67f1\u72b6\u306e\u7e2e\u5408\u7269\u304c\u89b3\u5bdf\u3055\u308c\u308b\u306e\u306b\u5bfe\u3057\u3001\u76f4\u9396\u72b6\u30d7\u30e9\u30b9\u30df\u30c9\u3067\u306f\u4f38\u9577\u3057\u305f\u30d1\u30fc\u30eb\u30cd\u30c3\u30af\u30ec\u30b9\u306e\u3088\u3046\u306a\u69cb\u9020\u304c\u89b3\u5bdf\u3055\u308c\u307e\u3059\u3002\u3088\u308a\u30b3\u30f3\u30d1\u30af\u30c8\u306a\u30b5\u30a4\u30ba\u306e\u74b0\u72b6\u30d7\u30e9\u30b9\u30df\u30c9\u306e\u30ab\u30c1\u30aa\u30f3\u6027\u8102\u8cea\u5a92\u4ecb\u6027\u30c8\u30e9\u30f3\u30b9\u30d5\u30a7\u30af\u30b7\u30e7\u30f3\u306f\u30a8\u30f3\u30c9\u30b5\u30a4\u30c8\u30fc\u30b7\u30b9\u7d4c\u8def\u306b\u4fb5\u5165\u3059\u308b\u53ef\u80fd\u6027\u304c\u3042\u308a\u307e\u3059\u304c\u3001\u4f38\u9577\u3057\u305f\u76f4\u9396\u72b6DNA\u69cb\u9020\u3067\u306f\u672c\u7d4c\u8def\u3078\u306e\u4fb5\u5165\u306f\u305d\u308c\u3068\u306f\u304b\u306a\u308a\u7570\u306a\u308a\u3001\u52b9\u7387\u306f\u4f4e\u3044\u3068\u8003\u3048\u3089\u308c\u307e\u3059\uff08von Groll et al., 2006\uff09\u3002<\/p>\n

\u5b89\u5b9a\u30c8\u30e9\u30f3\u30b9\u30d5\u30a7\u30af\u30b7\u30e7\u30f3\u306f\u3001\u76f4\u9396\u72b6DNA\u3092\u4f7f\u7528\u3057\u305f\u5834\u5408\u306b\u5bbf\u4e3b\u30b2\u30ce\u30e0\u3078\u306e\u6700\u9069\u306a\u7d44\u307f\u8fbc\u307f\u304c\u751f\u3058\u308b\u305f\u3081\u3001\u3088\u308a\u52b9\u7387\u7684\u3067\u3059\u3002<\/strong>\u30d5\u30ea\u30fc\u306e\u672b\u7aef\u3092\u6301\u3064\u76f4\u9396\u72b6DNA\u306f\u3001\u3088\u308a\u7d44\u63db\u3048\u304c\u8a98\u5c0e\u3055\u308c\u3001\u5bbf\u4e3b\u67d3\u8272\u4f53\u306b\u7d44\u307f\u8fbc\u307e\u308c\u3084\u3059\u304f\u3001\u7d30\u80de\u3078\u306e\u53d6\u308a\u8fbc\u307f\u52b9\u7387\u304c\u4f4e\u304f\u3066\u3082\u5b89\u5b9a\u306a\u5f62\u8cea\u8ee2\u63db\u4f53\u3092\u751f\u3058\u307e\u3059\u3002<\/p>\n

\u30ab\u30c1\u30aa\u30f3\u6027\u8102\u8cea\u5a92\u4ecb\u6027\u30c8\u30e9\u30f3\u30b9\u30d5\u30a7\u30af\u30b7\u30e7\u30f3\u306b\u304a\u3051\u308b\u53d6\u308a\u8fbc\u307f\u52b9\u7387\u304c\u540c\u7a0b\u5ea6\u3067\u3042\u3063\u3066\u3082\u3001\u30b5\u30a4\u30ba\u304c\u5927\u304d\u3044\u30d7\u30e9\u30b9\u30df\u30c9\u306e\u6838\u3078\u306e\u5c0e\u5165\u52b9\u7387\u306f\u5c0f\u3055\u3044\u30d7\u30e9\u30b9\u30df\u30c9\u3088\u308a\u3082\u4f4e\u4e0b\u3057\u307e\u3059\u3002<\/strong>\u3053\u306e\u5f71\u97ff\u306f\u3001\u8cea\u91cf\u307e\u305f\u306f\u30e2\u30eb\u6fc3\u5ea6\u304c\u7b49\u3057\u304f\u3001\u30b5\u30a4\u30ba\u304c\u7570\u306a\u308b\u30b3\u30f3\u30b9\u30c8\u30e9\u30af\u30c8\u3092\u7528\u3044\u308b\u3068\u89b3\u5bdf\u3055\u308c\u3001\u30d7\u30e9\u30b9\u30df\u30c9\u306e\u6838\u3078\u306e\u5c0e\u5165\u306f\u7d30\u80de\u5185\u79fb\u884c\u901f\u5ea6\u306b\u3088\u3063\u3066\u5236\u9650\u3055\u308c\u3001\u5c0f\u3055\u3044\u30d7\u30e9\u30b9\u30df\u30c9\u306f\u9ad8\u901f\u3067\u7d30\u80de\u8cea\u5185\u306b\u79fb\u884c\u3059\u308b\u3053\u3068\u306b\u3088\u308a\u5206\u89e3\u3055\u308c\u308b\u306e\u3092\u56de\u907f\u3057\u3066\u3044\u308b\u3068\u8003\u3048\u3089\u308c\u307e\u3059\uff08Lukacs, et al., 2000; McLenachan et al., 2007\uff09\u3002<\/p>\n

\u30d7\u30e9\u30b9\u30df\u30c9DNA\u306e\u54c1\u8cea<\/h3>\n

\u30d7\u30e9\u30b9\u30df\u30c9DNA\u306e\u7d14\u5ea6\u304a\u3088\u3073\u54c1\u8cea\u306f\u30c8\u30e9\u30f3\u30b9\u30d5\u30a7\u30af\u30b7\u30e7\u30f3\u3092\u6210\u529f\u3055\u305b\u308b\u305f\u3081\u306b\u91cd\u8981\u3067\u3059\u3002\u6700\u9ad8\u306e\u7d50\u679c\u3092\u9054\u6210\u3059\u308b\u305f\u3081\u306b\u306f\u3001\u30d5\u30a7\u30ce\u30fc\u30eb\u3001\u5869\u5316\u30ca\u30c8\u30ea\u30a6\u30e0\u3001\u304a\u3088\u3073\u30a8\u30f3\u30c9\u30c8\u30ad\u30b7\u30f3\u3092\u542b\u307e\u306a\u3044\u6700\u9ad8\u7d14\u5ea6\u306e\u30d7\u30e9\u30b9\u30df\u30c9DNA\u304c\u5fc5\u8981\u3067\u3059\u3002<\/strong>\u593e\u96d1\u7269\u306f\u7d30\u80de\u3092\u6b7b\u6ec5\u3055\u305b\u3001\u5869\u306f\u8102\u8cea\u8907\u5408\u4f53\u306e\u5f62\u6210\u3092\u963b\u5bb3\u3057\u3001\u30c8\u30e9\u30f3\u30b9\u30d5\u30a7\u30af\u30b7\u30e7\u30f3\u52b9\u7387\u3092\u4f4e\u4e0b\u3055\u305b\u307e\u3059\u3002\u30a8\u30f3\u30c9\u30c8\u30ad\u30b7\u30f3\u306f\u30ea\u30dd\u591a\u7cd6\u3068\u3057\u3066\u3082\u77e5\u3089\u308c\u3001\u30d7\u30e9\u30b9\u30df\u30c9\u8abf\u88fd\u306e\u6eb6\u89e3\u30b9\u30c6\u30c3\u30d7\u4e2d\u306b\u653e\u51fa\u3055\u308c\u3001\u591a\u304f\u306e\u5834\u5408\u3001\u30d7\u30e9\u30b9\u30df\u30c9DNA\u3068\u3068\u3082\u306b\u5171\u7cbe\u88fd\u3055\u308c\u307e\u3059\u3002\u3053\u308c\u3089\u306e\u5b58\u5728\u306f\u3001\u521d\u4ee3\u7d30\u80de\u304a\u3088\u3073\u4ed6\u306e\u30bb\u30f3\u30b7\u30c6\u30a3\u30d6\u306a\u7d30\u80de\u306b\u304a\u3051\u308b\u30c8\u30e9\u30f3\u30b9\u30d5\u30a7\u30af\u30b7\u30e7\u30f3\u52b9\u7387\u3092\u660e\u3089\u304b\u306b\u4f4e\u4e0b\u3055\u305b\u307e\u3059\u3002DNA\u306e\u5206\u96e2\u306b\u306f\u3001\u6700\u9ad8\u54c1\u8cea\u306eDNA\u306e\u30c8\u30e9\u30f3\u30b9\u30d5\u30a7\u30af\u30b7\u30e7\u30f3\u3092\u63d0\u4f9b\u3059\u308b\u5f0a\u793e\u306ePureLink\u2122 HiPure Plasmid Kit \uff08Mini\u3001Midi\u3001Maxi\u3001Mega\u3001\u304a\u3088\u3073Giga\uff09\u306e\u4f7f\u7528\u3092\u304a\u85a6\u3081\u3057\u307e\u3059\u3002\u5869\u5316\u30bb\u30b7\u30a6\u30e0\u5bc6\u5ea6\u52fe\u914d\u9060\u5fc3\u6cd5\u3067\u751f\u3058\u305f\u30d0\u30f3\u30c9\u3092\u5206\u96e2\u3059\u308b\u3053\u3068\u306b\u3088\u3063\u3066\u3082\u9ad8\u7d14\u5ea6DNA\u304c\u5f97\u3089\u308c\u307e\u3059\u304c\u3001\u3053\u306e\u30d7\u30ed\u30bb\u30b9\u306f\u6642\u9593\u3092\u6d6a\u8cbb\u3057\u307e\u3059\u3002DNA-\u8102\u8cea\u8907\u5408\u4f53\u3084DNA\u6eb6\u6db2\u3092\u904e\u5270\u306b\u30dc\u30eb\u30c6\u30c3\u30af\u30b9\u3059\u308b\u3053\u3068\u306f\u3001\u7279\u306b\u5927\u304d\u3044\u5206\u5b50\u3092\u90e8\u5206\u7684\u306b\u526a\u65ad\u3059\u308b\u3053\u3068\u306b\u3064\u306a\u304c\u308a\u3001\u305d\u308c\u306b\u3088\u3063\u3066\u30c8\u30e9\u30f3\u30b9\u30d5\u30a7\u30af\u30b7\u30e7\u30f3\u52b9\u7387\u304c\u4f4e\u4e0b\u3057\u307e\u3059\u3002\u5e0c\u91c8\u6e08\u307f\u306eDNA\u306b\u304a\u3051\u308bEDTA\u6fc3\u5ea6\u306f0.3 mM\u3092\u8d85\u3048\u306a\u3044\u3088\u3046\u306b\u3057\u307e\u3059\u3002<\/p>\n

\u907a\u4f1d\u5b50\u7523\u7269\u304a\u3088\u3073\u30d7\u30ed\u30e2\u30fc\u30bf\u30fc<\/h3>\n

\u30d7\u30ed\u30e2\u30fc\u30bf\u30fc\u306e\u9078\u629e\u306f\u5bbf\u4e3b\u7d30\u80de\u7cfb\u3001\u904e\u5270\u767a\u73fe\u3055\u305b\u308b\u30bf\u30f3\u30d1\u30af\u8cea\u3001\u304a\u3088\u3073\u5e0c\u671b\u3059\u308b\u767a\u73fe\u30ec\u30d9\u30eb\u306b\u5fdc\u3058\u3066\u884c\u3044\u307e\u3059\u3002<\/strong>\u5e45\u5e83\u3044\u7d30\u80de\u30bf\u30a4\u30d7\u3067\u6700\u3082\u9ad8\u3044\u767a\u73fe\u6d3b\u6027\u3092\u793a\u3059\u5f37\u529b\u306aCMV\uff08\u30b5\u30a4\u30c8\u30e1\u30ac\u30ed\u30a6\u30a4\u30eb\u30b9\uff09\u30d7\u30ed\u30e2\u30fc\u30bf\u30fc\u306f\u591a\u304f\u306e\u7814\u7a76\u8005\u306b\u7528\u3044\u3089\u308c\u3066\u3044\u307e\u3059\u3002\u54fa\u4e73\u985e\u7d30\u80de\u3067\u30bf\u30f3\u30d1\u30af\u8cea\u3092\u9ad8\u767a\u73fe\u3055\u305b\u308b\u305f\u3081\u306e\u4ed6\u306e\u5f37\u529b\u306a\u30d7\u30ed\u30e2\u30fc\u30bf\u30fc\u3068\u3057\u3066EF-1\u03b1\uff08human elongation factor-1\u03b1\uff09\u304c\u3042\u308a\u307e\u3059\u3002\u3057\u304b\u3057\u3001\u5f37\u529b\u904e\u304e\u308b\u30d7\u30ed\u30e2\u30fc\u30bf\u30fc\u3092\u4f7f\u7528\u3059\u308b\u3053\u3068\u306f\u3001\u6bd2\u6027\u907a\u4f1d\u5b50\u767a\u73fe\u306e\u53ef\u80fd\u6027\u3092\u9ad8\u3081\u308b\u3053\u3068\u306b\u306a\u308b\u305f\u3081\u3001\u30d7\u30e9\u30b9\u30df\u30c9DNA\u306e\u4e00\u904e\u6027\u30c8\u30e9\u30f3\u30b9\u30d5\u30a7\u30af\u30b7\u30e7\u30f3\u306b\u304a\u3044\u3066\u554f\u984c\u304c\u3042\u308a\u307e\u3059\u3002\u6bd2\u6027\u907a\u4f1d\u5b50\u7523\u7269\u767a\u73fe\u306e\u53ef\u80fd\u6027\u304c\u3042\u308b\u5834\u5408\u306f\u3001\u5f31\u3044\u30d7\u30ed\u30e2\u30fc\u30bf\u30fc\u306e\u4f7f\u7528\u3092\u304a\u85a6\u3081\u3057\u307e\u3059\u3002<\/p>\n

\u6bd2\u6027\u907a\u4f1d\u5b50\u7523\u7269\u306f\u3001\u5b89\u5b9a\u7684\u306b\u30c8\u30e9\u30f3\u30b9\u30d5\u30a7\u30af\u30b7\u30e7\u30f3\u3055\u308c\u305f\u7d30\u80de\u3092\u9078\u629e\u3059\u308b\u969b\u306b\u3082\u554f\u984c\u3067\u3059\u3002\u6297\u751f\u7269\u8cea\u8010\u6027\u907a\u4f1d\u5b50\u3092\u767a\u73fe\u3059\u308b\u7d30\u80de\u306f\u3001\u305d\u306e\u907a\u4f1d\u5b50\u306e\u767a\u73fe\u306b\u3088\u3063\u3066\u5c0e\u5165\u3055\u308c\u305f\u7d30\u80de\u306e\u5065\u5168\u6027\u304c\u640d\u306a\u308f\u308c\u305f\u6642\u306b\u5897\u6b96\u512a\u4f4d\u6027\u3092\u5931\u3044\u307e\u3059\u3002\u305d\u308c\u306b\u3088\u3063\u3066\u3001\u69cb\u6210\u7684\u30d7\u30ed\u30e2\u30fc\u30bf\u30fc\u3092\u7528\u3044\u3066\u5b89\u5b9a\u7684\u306b\u30c8\u30e9\u30f3\u30b9\u30d5\u30a7\u30af\u30b7\u30e7\u30f3\u3055\u308c\u305f\u30af\u30ed\u30fc\u30f3\u3092\u7372\u5f97\u3059\u308b\u3053\u3068\u304c\u4e0d\u53ef\u80fd\u3067\u3059\u3002\u305d\u306e\u3088\u3046\u306a\u30b1\u30fc\u30b9\u3067\u306f\u3001\u907a\u4f1d\u5b50\u767a\u73fe\u306e\u30bf\u30a4\u30df\u30f3\u30b0\u3092\u30b3\u30f3\u30c8\u30ed\u30fc\u30eb\u3059\u308b\u305f\u3081\u306b\u8a98\u5c0e\u6027\u30d7\u30ed\u30e2\u30fc\u30bf\u30fc\u3092\u7528\u3044\u3066\u3001\u5b89\u5b9a\u306a\u5f62\u8cea\u79fb\u5165\u4f53\u3092\u9078\u629e\u3059\u308b\u3053\u3068\u304c\u53ef\u80fd\u3067\u3059\u3002\u8a98\u5c0e\u6027\u30d7\u30ed\u30e2\u30fc\u30bf\u30fc\u306f\u3001\u901a\u5e38\u3001\u6a5f\u80fd\u3059\u308b\u305f\u3081\u306b\u30a4\u30f3\u30c7\u30e5\u30fc\u30b5\u30fc\u5206\u5b50\uff08\u4f8b\uff1a \u91d1\u5c5e\u30a4\u30aa\u30f3\u3001\u4ee3\u8b1d\u7523\u7269\u3001\u307e\u305f\u306f\u30db\u30eb\u30e2\u30f3\uff09\u306e\u5b58\u5728\u3092\u5fc5\u8981\u3068\u3057\u307e\u3059\u304c\u3001\u4e00\u90e8\u306e\u30a4\u30f3\u30c7\u30e5\u30fc\u30b5\u30fc\u5206\u5b50\u306f\u9006\u306e\u65b9\u6cd5\u3001\u3059\u306a\u308f\u3061\u7279\u5b9a\u306e\u5206\u5b50\u306e\u975e\u5b58\u5728\u4e0b\u3067\u907a\u4f1d\u5b50\u767a\u73fe\u306e\u8a98\u5c0e\u304c\u6a5f\u80fd\u3057\u307e\u3059\u3002<\/p>\n

\u30b3\u30f3\u30c8\u30ed\u30fc\u30eb<\/h3>\n

\u30c8\u30e9\u30f3\u30b9\u30d5\u30a7\u30af\u30b7\u30e7\u30f3\u306e\u65b9\u6cd5\u306b\u95a2\u308f\u3089\u305a\u3001\u7d30\u80de\u306e\u5065\u5168\u6027\u3092\u78ba\u8a8d\u3059\u308b\u305f\u3081\u306e\u30b3\u30f3\u30c8\u30ed\u30fc\u30eb\u30c8\u30e9\u30f3\u30b9\u30d5\u30a7\u30af\u30b7\u30e7\u30f3\u3092\u5b9f\u65bd\u3059\u308b\u3053\u3068\u3001\u5831\u544a\u3055\u308c\u3066\u3044\u308b\u30a2\u30c3\u30bb\u30a4\u304c\u9069\u5207\u306b\u6a5f\u80fd\u3057\u3066\u3044\u308b\u304b\u3092\u6c7a\u5b9a\u3059\u308b\u3053\u3068\u3001\u304a\u3088\u3073\u3042\u3089\u3086\u308b\u633f\u5165\u95a2\u9023\u306e\u554f\u984c\u306e\u539f\u56e0\u3092\u660e\u3089\u304b\u306b\u3059\u308b\u3053\u3068\u306f\u91cd\u8981\u3067\u3059\u3002<\/strong>\u6700\u9069\u306a\u7d30\u80de\u5897\u6b96\u6761\u4ef6\u3092\u78ba\u8a8d\u3059\u308b\u305f\u3081\u306b\u306f\u3001\u30cd\u30ac\u30c6\u30a3\u30d6\u30b3\u30f3\u30c8\u30ed\u30fc\u30eb\uff08\u975eDNA\u3001\u975e\u30c8\u30e9\u30f3\u30b9\u30d5\u30a7\u30af\u30b7\u30e7\u30f3\u8a66\u85ac\uff09\u3092\u542b\u3080\u5fc5\u8981\u304c\u3042\u308a\u307e\u3059\u3002\u30ec\u30dd\u30fc\u30bf\u30fc\u30a2\u30c3\u30bb\u30a4\u304c\u9069\u5207\u306b\u6a5f\u80fd\u3059\u308b\u3053\u3068\u3092\u78ba\u7acb\u3059\u308b\u305f\u3081\u306b\u306f\u3001\u30dd\u30b8\u30c6\u30a3\u30d6\u30b3\u30f3\u30c8\u30ed\u30fc\u30eb\uff08\u78ba\u7acb\u3055\u308c\u305f\u30c8\u30e9\u30f3\u30b9\u30d5\u30a7\u30af\u30b7\u30e7\u30f3\u6cd5\u3067\u306e\u540c\u6642\u4e26\u884c\u7684\u30c8\u30e9\u30f3\u30b9\u30d5\u30a7\u30af\u30b7\u30e7\u30f3\uff09\u3092\u542b\u3080\u5fc5\u8981\u304c\u3042\u308a\u307e\u3059\u3002\u633f\u5165\u95a2\u9023\u306e\u554f\u984c\u304c\u5b58\u5728\u3059\u308b\u304b\u3092\u6c7a\u5b9a\u3059\u308b\u305f\u3081\u306b\u306f\u3001\u76ee\u7684\u306e\u907a\u4f1d\u5b50\u3092\u542b\u307e\u306a\u3044\u30d7\u30e9\u30b9\u30df\u30c9\u3092\u5c0e\u5165\u3059\u308b\u5fc5\u8981\u304c\u3042\u308a\u307e\u3059\u3002<\/p>\n

\u30d7\u30e9\u30b9\u30df\u30c9DNA\u30c8\u30e9\u30f3\u30b9\u30d5\u30a7\u30af\u30b7\u30e7\u30f3\u306e\u6700\u9069\u5316<\/h2>\n

\u3042\u3089\u3086\u308b\u30c8\u30e9\u30f3\u30b9\u30d5\u30a7\u30af\u30b7\u30e7\u30f3\u51e6\u7406\u306b\u304a\u3044\u3066\u4e0d\u53ef\u6b20\u306a\u6700\u521d\u306e\u30b9\u30c6\u30c3\u30d7\u306f\u30c8\u30e9\u30f3\u30b9\u30d5\u30a7\u30af\u30b7\u30e7\u30f3\u6761\u4ef6\u306e\u6700\u9069\u5316\u3067\u3059\u3002<\/strong>\u305d\u308c\u305e\u308c\u306e\u7d30\u80de\u30bf\u30a4\u30d7\u304a\u3088\u3073\u30c8\u30e9\u30f3\u30b9\u30d5\u30a7\u30af\u30b7\u30e7\u30f3\u51e6\u7406\u306b\u306f\u3001\u5916\u6765DNA\u306e\u6700\u9069\u306a\u5c0e\u5165\u306b\u5fc5\u8981\u3068\u3055\u308c\u308b\u7279\u5fb4\u7684\u306a\u4e00\u9023\u306e\u8981\u4ef6\u304c\u3042\u308a\u3001\u3053\u308c\u3089\u306e\u6761\u4ef6\u306f\u4e92\u3044\u306b\u975e\u5e38\u306b\u985e\u4f3c\u3057\u305f\u7d30\u80de\u30bf\u30a4\u30d7\u9593\u3067\u3082\u5927\u304d\u306a\u5909\u52d5\u304c\u3042\u308a\u307e\u3059\u3002<\/p>\n

\u30c8\u30e9\u30f3\u30b9\u30d5\u30a7\u30af\u30b7\u30e7\u30f3\u52b9\u7387\u3092\u6700\u9069\u5316\u3059\u308b\u306e\u306b\u6700\u3082\u91cd\u8981\u306a\u4e00\u3064\u306e\u8981\u7d20\u306f\u3001\u7d30\u80de\u30bf\u30a4\u30d7\u306b\u9069\u5207\u306a\u30c8\u30e9\u30f3\u30b9\u30d5\u30a7\u30af\u30b7\u30e7\u30f3\u30d7\u30ed\u30c8\u30b3\u30fc\u30eb\u3092\u9078\u629e\u3059\u308b\u3053\u3068\u3067\u3059\u3002<\/strong>\u4e00\u65e6\u9069\u5207\u306a\u30c8\u30e9\u30f3\u30b9\u30d5\u30a7\u30af\u30b7\u30e7\u30f3\u6cd5\u3092\u9078\u629e\u3057\u305f\u3089\u3001\u69d8\u3005\u306a\u6761\u4ef6\u4e0b\u3067\u30ec\u30dd\u30fc\u30bf\u30fc\u907a\u4f1d\u5b50\u3092\u5c0e\u5165\u3057\u3001\u30ec\u30dd\u30fc\u30bf\u30fc\u907a\u4f1d\u5b50\u7523\u7269\u306e\u30a2\u30c3\u30bb\u30a4\u3059\u308b\u3053\u3068\u306b\u3088\u308a\u30c8\u30e9\u30f3\u30b9\u30d5\u30a7\u30af\u30b7\u30e7\u30f3\u52b9\u7387\u3092\u30e2\u30cb\u30bf\u30fc\u3059\u308b\u3053\u3068\u306b\u3088\u3063\u3066\u4e00\u904e\u6027\u30ec\u30dd\u30fc\u30bf\u30fc\u30a2\u30c3\u30bb\u30a4\u30b7\u30b9\u30c6\u30e0\u3092\u7528\u3044\u3066\u624b\u9806\u3092\u6700\u9069\u5316\u3067\u304d\u307e\u3059\u3002<\/p>\n

\u3053\u3053\u3067\u306f\u3001Neon Transfection System<\/a>\u3092\u7528\u3044\u308b\u30a8\u30ec\u30af\u30c8\u30ed\u30dd\u30ec\u30fc\u30b7\u30e7\u30f3\u3001\u304a\u3088\u3073\u30ab\u30c1\u30aa\u30f3\u6027\u8102\u8cea\u5a92\u4ecb\u6027\u30c8\u30e9\u30f3\u30b9\u30d5\u30a7\u30af\u30b7\u30e7\u30f3\u306e\u6700\u9069\u5316\u306b\u95a2\u3059\u308b\u6709\u76ca\u306a\u60c5\u5831\u304a\u3088\u3073\u4e00\u822c\u7684\u306a\u30ac\u30a4\u30c9\u30e9\u30a4\u30f3\u3092\u3054\u7d39\u4ecb\u3057\u307e\u3059\u3002<\/p>\n

\u30ea\u30f3\u9178\u30ab\u30eb\u30b7\u30a6\u30e0\u5171\u6c88\u6bbf\u6cd5\u306b\u5f71\u97ff\u3059\u308b\u91cd\u8981\u306a\u8981\u7d20<\/h3>\n

\u30ea\u30f3\u9178\u30ab\u30eb\u30b7\u30a6\u30e0\u6cd5\u306b\u3088\u308b\u30c8\u30e9\u30f3\u30b9\u30d5\u30a7\u30af\u30b7\u30e7\u30f3\u52b9\u7387\u306b\u5f71\u97ff\u3092\u53ca\u307c\u3059\u4e3b\u8981\u306a\u8981\u7d20\u306f\u3001\u30ea\u30f3\u9178\u30ab\u30eb\u30b7\u30a6\u30e0-DNA\u5171\u6c88\u6bbf\u7269\u4e2d\u306b\u542b\u307e\u308c\u308bDNA\u91cf\u3001\u7d30\u80de\u3092\u5171\u6c88\u6bbf\u7269\u3068\u30a4\u30f3\u30ad\u30e5\u30d9\u30fc\u30b7\u30e7\u30f3\u3059\u308b\u6642\u9593\u306e\u9577\u3055\u3001\u306a\u3089\u3073\u306b\u30b0\u30ea\u30bb\u30ed\u30fc\u30eb\u30b7\u30e7\u30c3\u30af\u307e\u305f\u306fDMSO\u30b7\u30e7\u30c3\u30af\u306e\u4f7f\u7528\u304a\u3088\u3073\u51e6\u7406\u6642\u9593\u306e\u9577\u3055\u3067\u3059\u3002<\/strong><\/p>\n

\u30ea\u30f3\u9178\u30ab\u30eb\u30b7\u30a6\u30e0\u6cd5\u306b\u3088\u308b\u30c8\u30e9\u30f3\u30b9\u30d5\u30a7\u30af\u30b7\u30e7\u30f3\u306b\u304a\u3044\u3066\u7528\u3044\u3089\u308c\u308b\u30c8\u30fc\u30bf\u30ebDNA\u91cf\u306f\u3001\u901a\u5e38\u300110 cm\u30c7\u30a3\u30c3\u30b7\u30e51\u679a\u5f53\u305f\u308a\u3001450 \u03bcL\u306e\u6ec5\u83cc\u6c34\u306850 \u03bcL\u306e2.5 M CaCl2<\/sub>\u4e2d\u306b10\uff5e50 \u03bcg\u3067\u3001\u305d\u306e\u91cf\u306f\u30d7\u30e9\u30b9\u30df\u30c9\u8abf\u88fd\u6cd5\u306a\u3089\u3073\u306b\u7d30\u80de\u304a\u3088\u3073\u57f9\u5730\u306b\u3088\u3063\u3066\u5927\u304d\u304f\u7570\u306a\u308a\u307e\u3059\u300210\uff5e15 \u03bcg\u306eDNA\u309210 cm\u30c7\u30a3\u30c3\u30b7\u30e5\u306b\u6dfb\u52a0\u3059\u308b\u3068\u3001\u4e00\u90e8\u306e\u7d30\u80de\u7cfb\u3067\u306f\u904e\u5ea6\u306e\u7d30\u80de\u6b7b\u304c\u751f\u3058\u3001DNA\u304c\u307b\u3068\u3093\u3069\u53d6\u308a\u8fbc\u307e\u308c\u306a\u304b\u3063\u305f\u308a\u3001\u4ed6\u306e\u7d30\u80de\u7cfb\u3001\u7279\u306b\u521d\u4ee3\u7d30\u80de\u3067\u306f\u306f\u308b\u304b\u306b\u9ad8\u3044DNA\u6fc3\u5ea6\u304c\u5fc5\u8981\u3068\u3055\u308c\u305f\u308a\u307e\u3059\u3002\u3053\u306e\u305f\u3081\u3001\u65b0\u305f\u306b\u30d7\u30e9\u30b9\u30df\u30c9\u3092\u8abf\u88fd\u3059\u308b\u5ea6\u306b\u3001\u307e\u305f\u65b0\u3057\u3044\u7d30\u80de\u7cfb\u3092\u30c8\u30e9\u30f3\u30b9\u30d5\u30a7\u30af\u30b7\u30e7\u30f3\u306b\u7528\u3044\u308b\u5ea6\u306b\u6700\u9069\u306aDNA\u6fc3\u5ea6\u306b\u3064\u3044\u3066\u8a66\u9a13\u3092\u884c\u3046\u5fc5\u8981\u304c\u3042\u308a\u307e\u3059\u3002<\/p>\n

\u7d30\u80de\u3092\u5171\u6c88\u6bbf\u7269\u3068\u30a4\u30f3\u30ad\u30e5\u30d9\u30fc\u30b7\u30e7\u30f3\u3059\u308b\u6700\u9069\u306a\u6642\u9593\u306e\u9577\u3055\u3082\u3001\u7d30\u80de\u30bf\u30a4\u30d7\u306b\u3088\u3063\u3066\u7570\u306a\u308a\u307e\u3059\u3002HeLa\u3001NIH 3T3\u3001\u304a\u3088\u3073BALB\/c 3T3\u306a\u3069\u306e\u4e00\u90e8\u306e\u9811\u5f37\u306a\u7d30\u80de\u30bf\u30a4\u30d7\u306f\u3001\u6700\u592716\u6642\u9593\u3067\u5171\u6c88\u6bbf\u7269\u304b\u3089\u5206\u96e2\u3059\u308b\u3053\u3068\u306b\u3088\u308a\u52b9\u7387\u7684\u306b\u30c8\u30e9\u30f3\u30b9\u30d5\u30a7\u30af\u30b7\u30e7\u30f3\u3055\u308c\u307e\u3059\u3002<\/p>\n

DNA\u91cf\u3001\u30a4\u30f3\u30ad\u30e5\u30d9\u30fc\u30b7\u30e7\u30f3\u6642\u9593\u3001\u304a\u3088\u3073\u30b0\u30ea\u30bb\u30ed\u30fc\u30eb\u30b7\u30e7\u30c3\u30af\u307e\u305f\u306fDMSO\u30b7\u30e7\u30c3\u30af\u3078\u306e\u66dd\u9732\u3092\u5909\u5316\u3055\u305b\u308b\u30d1\u30a4\u30ed\u30c3\u30c8\u5b9f\u9a13\u3067\u306f\u3001\u30ea\u30f3\u9178\u30ab\u30eb\u30b7\u30a6\u30e0\u6c88\u6bbf\u3078\u306e\u9577\u6642\u9593\u66dd\u9732\u306b\u5bfe\u3059\u308b\u8010\u6027\u3092\u6301\u3064\u7d30\u80de\u30bf\u30a4\u30d7\u3001\u30b0\u30ea\u30bb\u30ed\u30fc\u30eb\u30b7\u30e7\u30c3\u30af\u3092\u4f7f\u7528\u3059\u3079\u304d\u7d30\u80de\u30bf\u30a4\u30d7\u3092\u660e\u3089\u304b\u306b\u3067\u304d\u307e\u3059\u3002\u30d1\u30a4\u30ed\u30c3\u30c8\u5b9f\u9a13\u306e\u7d50\u679c\u304c\u5f97\u3089\u308c\u305f\u3089\u3001\u5b9f\u9a13\u5909\u6570\u3092\u3055\u3089\u306b\u7d30\u304b\u304f\u8abf\u7bc0\u3059\u308b\u3053\u3068\u306b\u3088\u308a\u3055\u3089\u306a\u308b\u6700\u9069\u5316\u3092\u884c\u3048\u307e\u3059\u3002\u4f8b\u3048\u3070\u3001\u4e0b\u8868\u306e\u4f8b\u306e\u3088\u3046\u306b\u7d30\u80de\u30923\u5206\u959310%\u30b0\u30ea\u30bb\u30ed\u30fc\u30eb\u3067\u30b7\u30e7\u30c3\u30af\u51e6\u7406\u3059\u308b\u3068\u3001\u30c8\u30e9\u30f3\u30b9\u30d5\u30a7\u30af\u30b7\u30e7\u30f3\u52b9\u7387\u304c\u5897\u5f37\u3055\u308c\u307e\u3059\u3002\u3042\u308b\u3044\u306f\u3001\u30b0\u30ea\u30bb\u30ed\u30fc\u30eb\u30b7\u30e7\u30c3\u30af\u51e6\u7406\u6642\u9593\u3092\u5909\u5316\u3055\u305b\u305f\u308a\u300110\uff5e20%\u306eDMSO\u30b7\u30e7\u30c3\u30af\u3092\u7528\u3044\u308b\u5b9f\u9a13\u3092\u884c\u3046\u3053\u3068\u3082\u3042\u308a\u307e\u3059\u3002<\/p>\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n
\u8868 \u30ea\u30f3\u9178\u30ab\u30eb\u30b7\u30a6\u30e0\u5171\u6c88\u6bbf\u6cd5\u306b\u3088\u308b\u30c8\u30e9\u30f3\u30b9\u30d5\u30a7\u30af\u30b7\u30e7\u30f3\u306e\u6700\u9069\u5316\u306e\u305f\u3081\u306e\u30d1\u30a4\u30ed\u30c3\u30c8\u5b9f\u9a13\u306e\u4f8b<\/caption>\n
\u30c7\u30a3\u30c3\u30b7\u30e5
\n\uff0810 cm\uff09<\/td>\n		\u30ec\u30dd\u30fc\u30bf\u30fc\u30d7\u30e9\u30b9\u30df\u30c9
\n\uff08\u03bcg\uff09<\/td>\n		\u30a4\u30f3\u30ad\u30e5\u30d9\u30fc\u30b7\u30e7\u30f3
\n\uff08\u6642\u9593\uff09<\/td>\n		\u30b0\u30ea\u30bb\u30ed\u30fc\u30eb
\n\uff08\u5206\uff09<\/td>\n<\/tr>\n
1<\/td>\n5<\/td>\n6<\/td>\n–<\/td>\n<\/tr>\n
2<\/td>\n10<\/td>\n6<\/td>\n–<\/td>\n<\/tr>\n
3<\/td>\n15<\/td>\n6<\/td>\n–<\/td>\n<\/tr>\n
4<\/td>\n20<\/td>\n16<\/td>\n–<\/td>\n<\/tr>\n
5<\/td>\n25<\/td>\n16<\/td>\n–<\/td>\n<\/tr>\n
6<\/td>\n30<\/td>\n16<\/td>\n–<\/td>\n<\/tr>\n
7<\/td>\n5<\/td>\n6<\/td>\n3<\/td>\n<\/tr>\n
8<\/td>\n10<\/td>\n6<\/td>\n3<\/td>\n<\/tr>\n
9<\/td>\n15<\/td>\n6<\/td>\n3<\/td>\n<\/tr>\n
10<\/td>\n20<\/td>\n16<\/td>\n3<\/td>\n<\/tr>\n
11<\/td>\n25<\/td>\n16<\/td>\n3<\/td>\n<\/tr>\n
12<\/td>\n30<\/td>\n16<\/td>\n3<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

\u30ab\u30c1\u30aa\u30f3\u6027\u8102\u8cea\u5a92\u4ecb\u6027\u5c0e\u5165\u306b\u5f71\u97ff\u3059\u308b\u91cd\u8981\u306a\u8981\u7d20<\/h3>\n

\u30ab\u30c1\u30aa\u30f3\u6027\u30ea\u30dd\u30bd\u30fc\u30e0\u306b\u3088\u308bDNA\u30c8\u30e9\u30f3\u30b9\u30d5\u30a7\u30af\u30b7\u30e7\u30f3\u306e\u6210\u529f\u306b\u306f\u3001DNA\u91cf\u3001DNA\u306b\u5bfe\u3059\u308b\u30c8\u30e9\u30f3\u30b9\u30d5\u30a7\u30af\u30b7\u30e7\u30f3\u8a66\u85ac\u306e\u6bd4\u7387\u3001\u8102\u8cea-DNA\u8907\u5408\u4f53\u306e\u30a4\u30f3\u30ad\u30e5\u30d9\u30fc\u30b7\u30e7\u30f3\u6642\u9593\u3001\u304a\u3088\u3073\u8907\u5408\u4f53\u6dfb\u52a0\u6642\u306e\u7d30\u80de\u5bc6\u5ea6\u306e4\u3064\u306e\u4e3b\u8981\u306a\u30d1\u30e9\u30e1\u30fc\u30bf\u304c\u5f71\u97ff\u3057\u307e\u3059\u3002<\/strong>\u3053\u308c\u3089\u306e\u8981\u7d20\u306f\u3001\u5404\u7d30\u80de\u30bf\u30a4\u30d7\u304a\u3088\u3073\u30d9\u30af\u30bf\u30fc\u306e\u7d44\u307f\u5408\u308f\u305b\u306b\u3064\u3044\u3066\u4f53\u7cfb\u7684\u306b\u691c\u8a0e\u3059\u308b\u5fc5\u8981\u304c\u3042\u308a\u307e\u3059\u304c\u3001\u4e00\u65e6\u6700\u9069\u5316\u3059\u308b\u3068\u3001\u305d\u308c\u4ee5\u964d\u306e\u5168\u3066\u306e\u5b9f\u9a13\u306b\u304a\u3044\u3066\u4e00\u8cab\u6027\u304c\u7dad\u6301\u3055\u308c\u3001\u78ba\u5b9f\u306b\u518d\u73fe\u6027\u306e\u3042\u308b\u7d50\u679c\u304c\u5f97\u3089\u308c\u307e\u3059\u3002<\/p>\n

DNA\u91cf<\/h4>\n

DNA\u306e\u6700\u9069\u91cf\u306f\u3001\u5c0e\u5165\u3055\u308c\u305f\u30d7\u30e9\u30b9\u30df\u30c9\uff08\u4f8b\uff1a \u30d7\u30ed\u30e2\u30fc\u30bf\u30fc\u3001\u30d7\u30e9\u30b9\u30df\u30c9\u306e\u30b5\u30a4\u30ba\u3001\u8907\u88fd\u8d77\u70b9\uff09\u3001\u5c0e\u5165\u3055\u308c\u308b\u7d30\u80de\u306e\u6570\u3001\u57f9\u990a\u30c7\u30a3\u30c3\u30b7\u30e5\u306e\u30b5\u30a4\u30ba\u3001\u304a\u3088\u3073\u4f7f\u7528\u3055\u308c\u308b\u30bf\u30fc\u30b2\u30c3\u30c8\u7d30\u80de\u7cfb\u306b\u3088\u3063\u3066\u7570\u306a\u308a\u307e\u3059\u3002<\/strong>\u8a66\u9a13\u3092\u884c\u3063\u305f\u591a\u304f\u306e\u7d30\u80de\u30bf\u30a4\u30d7\u306b\u304a\u3044\u3066\u3001\u6bd4\u8f03\u7684\u5c11\u91cf\u306eDNA\u304c\u52b9\u7387\u7684\u306b\u53d6\u308a\u8fbc\u307e\u308c\u3001\u767a\u73fe\u3055\u308c\u3066\u3044\u307e\u3059\u3002\u5b9f\u969b\u3001\u4e00\u90e8\u306e\u7d30\u80de\u30bf\u30a4\u30d7\u3067\u306f\u3001\u9ad8\u30ec\u30d9\u30eb\u306eDNA\u91cf\u304c\u30ab\u30c1\u30aa\u30f3\u6027\u8102\u8cea\u8abf\u88fd\u7269\u306b\u5bfe\u3057\u3066\u963b\u5bb3\u6d3b\u6027\u3092\u793a\u3059\u53ef\u80fd\u6027\u304c\u3042\u308a\u307e\u3059\u3002\u307e\u305f\u3001\u6bd2\u6027\u30bf\u30f3\u30d1\u30af\u8cea\u3092\u30b3\u30fc\u30c9\u3059\u308b\u30d7\u30e9\u30b9\u30df\u30c9\u306e\u4f7f\u7528\u3084\u9ad8\u767a\u73fe\u7387\u3092\u3082\u305f\u3089\u3059\u904e\u5270\u306a\u30d7\u30e9\u30b9\u30df\u30c9\u3092\u4f7f\u7528\u3057\u305f\u5834\u5408\u3082\u7d30\u80de\u6bd2\u6027\u304c\u751f\u3058\u308b\u53ef\u80fd\u6027\u304c\u3042\u308a\u307e\u3059\u3002<\/p>\n

DNA\u306b\u5bfe\u3059\u308b\u30c8\u30e9\u30f3\u30b9\u30d5\u30a7\u30af\u30b7\u30e7\u30f3\u8a66\u85ac\u306e\u6bd4\u7387<\/h4>\n

\u30c8\u30e9\u30f3\u30b9\u30d5\u30a7\u30af\u30b7\u30e7\u30f3\u8907\u5408\u4f53\u306e\u5168\u4f53\u7684\u306a\u6b63\u5473\u306e\u96fb\u8377\u91cf\u306f\u3001DNA\u306b\u5bfe\u3059\u308b\u30c8\u30e9\u30f3\u30b9\u30d5\u30a7\u30af\u30b7\u30e7\u30f3\u8a66\u85ac\u306e\u6bd4\u7387\u306b\u3088\u3063\u3066\u6c7a\u5b9a\u3055\u308c\u307e\u3059\u3002<\/strong>DNA\u9aa8\u683c\u4e2d\u306e\u30ea\u30f3\u9178\u306b\u3088\u308b\u8ca0\u96fb\u8377\u306f\u3001\u826f\u597d\u306b\u8907\u5408\u4f53\u3092\u5f62\u6210\u3055\u305b\u308b\u305f\u3081\u306b\u3082\u3001\u8ca0\u306b\u8377\u96fb\u3057\u3066\u3044\u308b\u7d30\u80de\u819c\u7531\u6765\u306eDNA\u4e0a\u306e\u9759\u96fb\u53cd\u767a\u529b\u3092\u4e2d\u548c\u3059\u308b\u305f\u3081\u306b\u3082\u3001\u30c8\u30e9\u30f3\u30b9\u30d5\u30a7\u30af\u30b7\u30e7\u30f3\u8a66\u85ac\u7531\u6765\u306e\u6b63\u8377\u96fb\u306b\u3088\u3063\u3066\u76f8\u6bba\u3055\u305b\u308b\u5fc5\u8981\u304c\u3042\u308a\u307e\u3059\u3002<\/p>\n

DNA\u306b\u5bfe\u3059\u308b\u30c8\u30e9\u30f3\u30b9\u30d5\u30a7\u30af\u30b7\u30e7\u30f3\u8a66\u85ac\u306e\u6700\u9069\u306a\u6bd4\u7387\u306f\u3001\u7d30\u80de\u30bf\u30a4\u30d7\u306b\u5927\u304d\u304f\u4f9d\u5b58\u3057\u307e\u3059\u3002\u958b\u59cb\u6642\u70b9\u306b\u304a\u3044\u3066\u3001\u30d7\u30e9\u30b9\u30df\u30c9DNA\u6fc3\u5ea6\u306f\u4e00\u5b9a\u306b\u4fdd\u3061\u3001\u30c8\u30e9\u30f3\u30b9\u30d5\u30a7\u30af\u30b7\u30e7\u30f3\u8a66\u85ac\u306e\u91cf\u3092\u5909\u5316\u3055\u305b\u307e\u3059\uff08\u4f8b\uff1a 1\uff1a1\u30013\uff1a1\u304a\u3088\u30735\uff1a1 \u8cea\u91cf\uff0f\u4f53\u7a4d\u6bd4\uff09\u3002\u3053\u306e\u6bd4\u7387\u3092\u7dad\u6301\u3057\u3066\u3001\u6dfb\u52a0\u3059\u308b\u30d7\u30e9\u30b9\u30df\u30c9\u91cf\u3092\u5897\u52a0\u3055\u305b\u308b\u3053\u3068\u306b\u3088\u3063\u3066\u3001\u3055\u3089\u306a\u308b\u30e1\u30ea\u30c3\u30c8\u304c\u5f97\u3089\u308c\u308b\u53ef\u80fd\u6027\u304c\u3042\u308a\u307e\u3059\u3002<\/p>\n

\u30a4\u30f3\u30ad\u30e5\u30d9\u30fc\u30b7\u30e7\u30f3\u6642\u9593<\/h4>\n

\u7d30\u80de\u3068\u30c8\u30e9\u30f3\u30b9\u30d5\u30a7\u30af\u30b7\u30e7\u30f3\u8907\u5408\u4f53\u306e\u6700\u9069\u306a\u30a4\u30f3\u30ad\u30e5\u30d9\u30fc\u30b7\u30e7\u30f3\u6642\u9593\u306f\u7d30\u80de\u7cfb\u304a\u3088\u3073\u4f7f\u7528\u3057\u305f\u30c8\u30e9\u30f3\u30b9\u30d5\u30a7\u30af\u30b7\u30e7\u30f3\u8a66\u85ac\u306b\u4f9d\u5b58\u3057\u307e\u3059\u3002<\/strong>\u901a\u5e38\u3001\u30c8\u30e9\u30f3\u30b9\u30d5\u30a7\u30af\u30b7\u30e7\u30f3\u52b9\u7387\u306f\u8102\u8cea\u8a66\u85ac-DNA\u8907\u5408\u4f53\u3078\u306e\u66dd\u9732\u6642\u9593\u306b\u4f34\u3063\u3066\u5897\u52a0\u3057\u307e\u3059\u3002\u3068\u306f\u3044\u3048\u3001\u7279\u5b9a\u306e\u8102\u8cea\u8a66\u85ac\u3078\u306e\u9577\u6642\u9593\u66dd\u9732\u306f\u6bd2\u6027\u72b6\u614b\u3092\u9ad8\u3081\u308b\u305f\u3081\u3001\u30a4\u30f3\u30ad\u30e5\u30d9\u30fc\u30b7\u30e7\u30f3\u6642\u9593\u304c\u7d42\u4e86\u3057\u305f\u3089\u3001\u7d30\u80de\u6bd2\u6027\u306e\u5f71\u97ff\u3092\u6700\u5c0f\u9650\u306b\u6291\u3048\u308b\u305f\u3081\u306b\u3001\u9060\u5fc3\u5206\u96e2\u306b\u3088\u308b\u9664\u53bb\u3084\u65b0\u3057\u3044\u57f9\u5730\u306b\u3088\u308b\u5e0c\u91c8\u304c\u5fc5\u8981\u3068\u3055\u308c\u307e\u3059\u3002Lipofectamine 3000<\/a>\u8a66\u85ac\u306e\u3088\u3046\u306a\u65b0\u3057\u3044\u7a4f\u548c\u306a\u8a66\u85ac\u3067\u306f\u30c8\u30e9\u30f3\u30b9\u30d5\u30a7\u30af\u30b7\u30e7\u30f3\u5f8c\u306e\u8907\u5408\u4f53\u306e\u9664\u53bb\u3084\u5e0c\u91c8\u306f\u5fc5\u8981\u3042\u308a\u307e\u305b\u3093\u3002<\/p>\n

\u57f9\u5730\u306e\u6dfb\u52a0\u3084\u7f6e\u63db\u304c\u5fc5\u8981\u3068\u3055\u308c\u308b\u30ab\u30c1\u30aa\u30f3\u6027\u8102\u8cea\u8a66\u85ac\u3092\u4f7f\u7528\u3059\u308b\u5834\u5408\u306f\u3001\u8907\u5408\u4f53\u6dfb\u52a0\u5f8c\u306e\u30a4\u30f3\u30ad\u30e5\u30d9\u30fc\u30b7\u30e7\u30f3\u6642\u9593\u3092\u5909\u5316\u3055\u305b\u3066\uff08\u4f8b\uff1a 30\u5206\uff5e4\u6642\u9593\u3001\u307e\u305f\u306f\u4e00\u6669\uff09\u3001\u3053\u306e\u9593\u306e\u7d30\u80de\u5f62\u614b\u3092\u30e2\u30cb\u30bf\u30fc\u3057\u307e\u3059\u3002\u7279\u306b\u7d30\u80de\u3092\u7121\u8840\u6e05\u57f9\u5730\u306b\u7dad\u6301\u3057\u305f\u5834\u5408\u306f\u3001\u4e00\u90e8\u306e\u7d30\u80de\u7cfb\u3067\u306f\u3053\u308c\u3089\u306e\u6761\u4ef6\u4e0b\u3067\u751f\u5b58\u80fd\u304c\u5931\u308f\u308c\u307e\u3059\u3002<\/p>\n

\u7d30\u80de\u5bc6\u5ea6<\/h4>\n

\u7d30\u80de\u5bc6\u5ea6\u306f\u5168\u30c8\u30e9\u30f3\u30b9\u30d5\u30a7\u30af\u30b7\u30e7\u30f3\u52b9\u7387\u306b\u3082\u5f71\u97ff\u3057\u307e\u3059\u3002<\/strong>\u8ee2\u5199\u304a\u3088\u3073\u6700\u7d42\u7684\u306a\u30bf\u30f3\u30d1\u30af\u8cea\u751f\u7523\u3092\u9054\u6210\u3059\u308b\u305f\u3081\u306b\u306f\u3001DNA\u306e\u6838\u5185\u5c40\u5728\u304c\u5fc5\u8981\u3068\u3055\u308c\u3001\u305d\u308c\u306f\u6709\u7cf8\u5206\u88c2\u4e2d\u306e\u819c\u306e\u5206\u89e3\u3068\u518d\u7de8\u6210\u306b\u5927\u304d\u304f\u4f9d\u5b58\u3057\u3001\u7d30\u80de\u304c\u6d3b\u767a\u306b\u5206\u88c2\u3059\u308b\u3053\u3068\u304c\u5fc5\u8981\u3068\u3055\u308c\u307e\u3059\u3002<\/p>\n

\u63a5\u7740\u7d30\u80de\u3067\u306f\u3001\u591a\u304f\u306e\u5834\u5408\u300180%\u306e\u30b3\u30f3\u30d5\u30eb\u30a8\u30f3\u30b7\u30fc\u3067\u6700\u9ad8\u306e\u52b9\u7387\u306b\u5230\u9054\u3057\u307e\u3059\u304c\u3001\u30d7\u30ed\u30c8\u30b3\u30fc\u30eb\u3067\u306f40\uff5e90%\u306e\u7bc4\u56f2\u306e\u30b3\u30f3\u30d5\u30eb\u30a8\u30f3\u30b7\u30fc\u304c\u63a8\u5968\u3055\u308c\u3066\u3044\u307e\u3059\u3002\u6d6e\u904a\u7d30\u80de\u3067\u306f\u3001\u78ba\u5b9f\u306b\u30c8\u30e9\u30f3\u30b9\u30d5\u30a7\u30af\u30b7\u30e7\u30f3\u51e6\u7406\u306b\u6700\u9069\u306a\u751f\u7406\u7684\u72b6\u614b\u306b\u3057\u3066\u304a\u304f\u305f\u3081\u306b\u3001\u30c8\u30e9\u30f3\u30b9\u30d5\u30a7\u30af\u30b7\u30e7\u30f3\u306e1\u65e5\u524d\u306b\u7d30\u80de\u3092\u5206\u88c2\u3055\u305b\u308b\u3053\u3068\u3092\u63a8\u5968\u3057\u307e\u3059\u3002\u6700\u9069\u306a\u7d30\u80de\u5bc6\u5ea6\u306f\u3001\u7d30\u80de\u30bf\u30a4\u30d7\u304a\u3088\u3073\u8a66\u85ac\u7279\u7570\u7684\u306a\u6bd2\u6027\u306b\u5927\u304d\u304f\u4f9d\u5b58\u3059\u308b\u305f\u3081\u3001\u7d4c\u9a13\u7684\u306b\u6c7a\u5b9a\u3059\u308b\u5fc5\u8981\u304c\u3042\u308a\u307e\u3059\u3002<\/p>\n

\u30a8\u30ec\u30af\u30c8\u30ed\u30dd\u30ec\u30fc\u30b7\u30e7\u30f3\u306b\u5f71\u97ff\u3059\u308b\u91cd\u8981\u306a\u8981\u7d20<\/h3>\n

\u30a8\u30ec\u30af\u30c8\u30ed\u30dd\u30ec\u30fc\u30b7\u30e7\u30f3\u306f\u3001\u30d1\u30eb\u30b9\u96fb\u5727\u3001\u30d1\u30eb\u30b9\u5e45\u3001\u304a\u3088\u3073\u30d1\u30eb\u30b9\u6570\u306e\u4e3b\u306b3\u7a2e\u985e\u306e\u96fb\u6c17\u7684\u30d1\u30e9\u30e1\u30fc\u30bf\u306e\u7d44\u307f\u5408\u308f\u305b\u306b\u4f9d\u5b58\u3057\u307e\u3059\u3002<\/strong>\u30a8\u30ec\u30af\u30c8\u30ed\u30dd\u30ec\u30fc\u30b7\u30e7\u30f3\u306f\u5316\u5b66\u7684\u624b\u6cd5\u30d9\u30fc\u30b9\u306e\u30d7\u30ed\u30c8\u30b3\u30fc\u30eb\u3067\u306f\u306a\u3044\u305f\u3081\u3001DNA\u6fc3\u5ea6\u306e\u5f71\u97ff\u3092\u307b\u3068\u3093\u3069\u53d7\u3051\u307e\u305b\u3093\u3002\u3057\u304b\u3057\u3001\u30ea\u30f3\u9178\u30ab\u30eb\u30b7\u30a6\u30e0\u5a92\u4ecb\u6027\u30c8\u30e9\u30f3\u30b9\u30d5\u30a7\u30af\u30b7\u30e7\u30f3\u306e\u307b\u307c5\u500d\u4ee5\u4e0a\u306e\u7d30\u80de\u304a\u3088\u3073DNA\u304c\u5fc5\u8981\u3068\u3055\u308c\u307e\u3059\u3002\u901a\u5e38\u3001107<\/sup>\u500b\u306e\u7d30\u80de\u5f53\u305f\u308a1\uff5e5 \u03bcg\u306eDNA\u304c\u3042\u308c\u3070\u5341\u5206\u3067\u3001DNA\u306e\u5b58\u5728\u91cf\u3068\u53d6\u308a\u8fbc\u307e\u308c\u308b\u91cf\u3068\u306e\u9593\u306b\u9ad8\u3044\u76f8\u95a2\u304c\u3042\u308a\u307e\u3059\u3002<\/p>\n

\u30a8\u30ec\u30af\u30c8\u30ed\u30dd\u30ec\u30fc\u30b7\u30e7\u30f3\u30d1\u30e9\u30e1\u30fc\u30bf\u3092\u6700\u9069\u5316\u3059\u308b\u305f\u3081\u306b\u306f\u300140\uff5e80%\u306e\u7d30\u80de\u751f\u5b58\u7387\u3092\u7dad\u6301\u3059\u308b\u30d1\u30eb\u30b9\u3092\u898b\u3064\u3051\u307e\u3059\u3002\u30d1\u30eb\u30b9\u5e45\u306f\u3001\u96fb\u6e90\u306e\u96fb\u6c17\u5bb9\u91cf\u306b\u3088\u3063\u3066\u6c7a\u5b9a\u3055\u308c\u3001\u96fb\u6e90\u306b\u3088\u3063\u3066\u767a\u751f\u3059\u308b\u96fb\u5727\u306b\u4f9d\u5b58\u3057\u3066\u5909\u5316\u3057\u307e\u3059\u3002\u904e\u5ea6\u306e\u7d30\u80de\u6b7b\u304c\u751f\u3058\u308b\u5834\u5408\u306f\u3001\u96fb\u6c17\u5bb9\u91cf\u3092\u4f4e\u4e0b\u3055\u305b\u308b\u3053\u3068\u306b\u3088\u308a\u30d1\u30eb\u30b9\u306e\u9577\u3055\u3092\u77ed\u304f\u3067\u304d\u307e\u3059\u3002\u7d30\u80de\u3092\u6c37\u4e0a\u306b\u7dad\u6301\u3059\u308b\u3053\u3068\u3067\u3001\u591a\u304f\u306e\u5834\u5408\uff08\u7279\u306b\u71b1\u3092\u767a\u751f\u3059\u308b\u9ad8\u51fa\u529b\u96fb\u6e90\u3067\u306f\uff09\u3001\u7d30\u80de\u751f\u5b58\u7387\u304c\u6539\u5584\u3057\u3001\u3088\u308a\u52b9\u7387\u7684\u306a\u30c8\u30e9\u30f3\u30b9\u30d5\u30a7\u30af\u30b7\u30e7\u30f3\u7387\u304c\u5f97\u3089\u308c\u307e\u3059\uff08Potter et al., 1984\uff09\u3002\u3057\u304b\u3057\u3001\u4e00\u90e8\u306e\u7d30\u80de\u7cfb\u3067\u306f\u3001\u4f4e\u96fb\u5727\uff0f\u9ad8\u5bb9\u91cf\u306e\u6761\u4ef6\u4e0b\u3067\u3001\u5ba4\u6e29\u306b\u304a\u3044\u3066\u9ad8\u52b9\u7387\u306a\u30a8\u30ec\u30af\u30c8\u30ed\u30dd\u30ec\u30fc\u30b7\u30e7\u30f3\u304c\u884c\u3048\u307e\u3059\uff08Chu et al., 1991\uff09\u3002<\/p>\n

Neon Transfection System<\/a>\u306f\u300118-\u304a\u3088\u307324-\u30a6\u30a7\u30eb\u7528\u306b\u6700\u9069\u5316\u3055\u308c\u305f\u30d7\u30ed\u30c8\u30b3\u30fc\u30eb\u304c\u4e88\u3081\u30d7\u30ed\u30b0\u30e9\u30e0\u3055\u308c\u3066\u304a\u308a\u3001\u6570\u65e5\u4ee5\u5185\u306b\u591a\u6570\u306e\u63a5\u7740\u304a\u3088\u3073\u6d6e\u904a\u7d30\u80de\u7cfb\u7528\u306e\u96fb\u6c17\u7684\u30d1\u30e9\u30e1\u30fc\u30bf\u3092\u8fc5\u901f\u306b\u6700\u9069\u5316\u3067\u304d\u307e\u3059\u3002\u307e\u305f\u3001Neon Transfection System\u7528\u306e\u7d30\u80de\u7cfb\u7279\u7570\u7684\u306b\u6700\u9069\u5316\u3055\u308c\u305f\u30d7\u30ed\u30c8\u30b3\u30fc\u30eb\u3082\u30a6\u30a7\u30d6\u30b5\u30a4\u30c8<\/a>\u304b\u3089\u7c21\u5358\u306b\u30c0\u30a6\u30f3\u30ed\u30fc\u30c9\u3059\u308b\u3053\u3068\u304c\u3067\u304d\u3001\u591a\u304f\u306e\u901a\u5e38\u7528\u3044\u3089\u308c\u3066\u3044\u308b\u7d30\u80de\u30bf\u30a4\u30d7\u3078\u306e\u30c8\u30e9\u30f3\u30b9\u30d5\u30a7\u30af\u30b7\u30e7\u30f3\u52b9\u7387\u3092\u6700\u5927\u5316\u3067\u304d\u307e\u3059\u3002<\/p>\n

\u3010\u7121\u6599\u30c0\u30a6\u30f3\u30ed\u30fc\u30c9\u3011Gibco\u7d30\u80de\u57f9\u990a\u57fa\u790e\u30cf\u30f3\u30c9\u30d6\u30c3\u30af<\/h2>\n

\u7d30\u80de\u57f9\u990a\u306b\u95a2\u3059\u308b\u57fa\u790e\u60c5\u5831\u3092\u89e3\u8aac\u3057\u305f\u30cf\u30f3\u30c9\u30d6\u30c3\u30af\u3092\u3054\u7528\u610f\u3057\u3066\u3044\u307e\u3059\uff08\u65e5\u672c\u8a9e\u7248\u3001\u7d04100\u30da\u30fc\u30b8\uff09\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
\n\u30c0\u30a6\u30f3\u30ed\u30fc\u30c9<\/a><\/p>\n

 <\/p>\n

 <\/p>\n

\u7814\u7a76\u7528\u306b\u306e\u307f\u4f7f\u7528\u3067\u304d\u307e\u3059\u3002\u8a3a\u65ad\u76ee\u7684\u304a\u3088\u3073\u305d\u306e\u624b\u7d9a\u304d\u4e0a\u3067\u306e\u4f7f\u7528\u306f\u3067\u304d\u307e\u305b\u3093\u3002<\/p>\n","protected":false},"excerpt":{"rendered":"

\u53e4\u5178\u7684\u306a\u30c8\u30e9\u30f3\u30b9\u30d5\u30a7\u30af\u30b7\u30e7\u30f3\u6280\u8853\u306f\u3001\u5f53\u521d\u3001\u30d7\u30e9\u30b9\u30df\u30c9DNA\u3092\u7d30\u80de\u306b\u5c0e\u5165\u3059\u308b\u305f\u3081\u306b\u958b\u767a\u3055\u308c\u307e\u3057\u305f\u304c\u3001\u30d7\u30e9\u30b9\u30df\u30c9DNA\u306f\u30c8\u30e9\u30f3\u30b9\u30d5\u30a7\u30af\u30b7\u30e7\u30f3\u306e\u305f\u3081\u306e\u6700\u3082\u4e00\u822c\u7684\u306a\u30d9\u30af\u30bf\u30fc\u3068\u3057\u3066\u73fe\u5728\u3082\u5b58\u5728\u3057\u3066\u3044\u307e\u3059\u3002\u4eca\u56de\u306f\u3001\u30d7\u30e9\u30b9\u30df\u30c9DNA\u3092\u7528\u3044\u305f\u30c8\u30e9\u30f3\u30b9\u30d5\u30a7\u30af\u30b7\u30e7\u30f3\u306b\u3064\u3044\u3066\u306e\u30ac\u30a4\u30c9\u30e9\u30a4\u30f3\u3068\u30d1\u30e9\u30e1\u30fc\u30bf\u306e\u6700\u9069\u5316\u65b9\u6cd5\u306b\u3064\u3044\u3066\u3054\u7d39\u4ecb\u3057\u307e\u3059\u3002<\/p>\n","protected":false},"author":988,"featured_media":2933,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"_acf_changed":false,"_kad_blocks_custom_css":"","_kad_blocks_head_custom_js":"","_kad_blocks_body_custom_js":"","_kad_blocks_footer_custom_js":"","_monsterinsights_skip_tracking":false,"_genesis_hide_title":false,"_genesis_hide_breadcrumbs":false,"_genesis_hide_singular_image":false,"_genesis_hide_footer_widgets":false,"_genesis_custom_body_class":"","_genesis_custom_post_class":"","_genesis_layout":"","_jetpack_newsletter_access":"","_jetpack_dont_email_post_to_subs":false,"_jetpack_newsletter_tier_id":0,"_jetpack_memberships_contains_paywalled_content":false,"_jetpack_feature_clip_id":0,"_jetpack_memberships_contains_paid_content":false,"footnotes":"","jetpack_post_was_ever_published":false},"categories":[159],"tags":[67],"division":[],"class_list":{"0":"post-569","1":"post","2":"type-post","3":"status-publish","4":"format-standard","5":"has-post-thumbnail","7":"category-cell","8":"tag-transfection","9":"entry"},"_selected_authors":"","_selected_reviewers":"","acf":[],"yoast_head":"\n\u30d7\u30e9\u30b9\u30df\u30c9DNA\u30c8\u30e9\u30f3\u30b9\u30d5\u30a7\u30af\u30b7\u30e7\u30f3\u306e\u305f\u3081\u306e\u30ac\u30a4\u30c9\u30e9\u30a4\u30f3\u3068\u6700\u9069\u5316\u65b9\u6cd5 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