{"id":2418,"date":"2013-03-21T14:05:11","date_gmt":"2013-03-21T05:05:11","guid":{"rendered":"http:\/\/dna.brc.riken.jp:8002\/?page_id=2418"},"modified":"2023-06-13T09:29:39","modified_gmt":"2023-06-13T00:29:39","slug":"technicalnotes12","status":"publish","type":"page","link":"http:\/\/dna.brc.riken.jp\/ja\/manual\/technicalnotes12","title":{"rendered":"PCR\u306e\u8a71\u3057"},"content":{"rendered":"<h3>PCR\u306e\u8a71\u3057 (1)<\/h3>\n<ul style=\"line-height: 1.5em;\">\n<li>\u5e02\u8ca9\u306ePCR\u7528\u8a66\u85ac\u306e\u5897\u5e45\u52b9\u7387\u304c\u683c\u6bb5\u306b\u4e0a\u304c\u3063\u3066\u3044\u308b\u3068\u306f\u3044\u3048\u3001PCR\u3067\u76ee\u7684\u306e\u30d0\u30f3\u30c9\u304c\u73fe\u308c\u306a\u3044\u3053\u3068\u306f\u6642\u3005\u7d4c\u9a13\u3057\u307e\u3059\u3002\u30c6\u30f3\u30d7\u30ec\u30fc\u30c8\u304c\u30b2\u30ce\u30e0DNA\u3060\u3063\u305f\u308a\u3001\u5897\u5e45\u9818\u57df\u304c\u9577\u304b\u3063\u305f\u308a\u3001GC\u30ea\u30c3\u30c1\u306e\u914d\u5217\u304c\u542b\u307e\u308c\u308b\u5834\u5408\u306f\u6761\u4ef6\u8a2d\u5b9a\u304c\u7279\u306b\u96e3\u3057\u304f\u306a\u308a\u307e\u3059\u3002\u6700\u8fd1\u3001\u30b2\u30ce\u30df\u30c3\u30afPCR\u3092\u884c\u3046\u6a5f\u4f1a\u304c\u591a\u304f\u306a\u308a\u3001\u5931\u6557\u3068\u6210\u529f\u306e\u4f53\u9a13\u304b\u3089\u5e7e\u3064\u304b\u306e\u6559\u8a13\u3092\u5f97\u307e\u3057\u305f\u3002<\/li>\n<li>\u30c6\u30f3\u30d7\u30ec\u30fc\u30c8\u306e\u6761\u4ef6\uff1a<br \/>\n\u30b2\u30ce\u30e0DNA\u62bd\u51fa\u306b\u306f\u69d8\u3005\u306a\u30d7\u30ed\u30c8\u30b3\u30fc\u30eb\u304c\u3042\u308a\u307e\u3059\u304c\u3001\u7279\u306b\u3053\u3060\u308f\u3063\u3066\u3044\u307e\u305b\u3093\u3002\u30d5\u30a7\u30ce\u30fc\u30eb\uff0d\u30af\u30ed\u30ed\u30db\u30eb\u30e0\u62bd\u51fa\u3092\u4e00\u56de\u3068\u30a8\u30bf\u30ce\u30fc\u30eb\u6c88\u6fb1\u3067\u7cbe\u88fd\u3059\u308b\u304b\u3001\u307e\u305f\u306f\u5e02\u8ca9\u306e\u30b2\u30ce\u30e0DNA\u62bd\u51fa\u30ab\u30e9\u30e0\u3067\u7cbe\u88fd\u3057\u3066\u3044\u307e\u3059\u3002PCR\u7528\u306e\u30c6\u30f3\u30d7\u30ec\u30fc\u30c8\u306e\u5834\u5408\u3001\u30b5\u30b6\u30f3\u30d6\u30ed\u30c3\u30c6\u30a3\u30f3\u30b0\u7528\u306e\u30b2\u30ce\u30e0DNA\u306e\u3088\u3046\u306b\u6570\u5341kb\u306e\u9577\u3055\u3092\u4fdd\u3064\u5fc5\u8981\u304c\u306a\u304f\u3001\u3080\u3057\u308d\u30dc\u30eb\u30c6\u30af\u30b9\u3067\u9069\u5ea6\u306b\u65ad\u7247\u5316\u3055\u305b\u305f\u65b9\u304c\u5897\u5e45\u3057\u3084\u3059\u3044\u611f\u304c\u3042\u308a\u307e\u3059\u3002RNA\u306e\u5927\u91cf\u6df7\u5165\u306fPCR\u53cd\u5fdc\u3092\u963b\u5bb3\u3059\u308b\u8981\u56e0\u306b\u306a\u308a\u3048\u307e\u3059\u304c\u3001DNA\u62bd\u51fa\u306e\u6700\u521d\u306e\u30b9\u30c6\u30c3\u30d7\u3067\u7d44\u7e54\u3092\u6eb6\u89e3\u3059\u308b\u30d0\u30c3\u30d5\u30a1\u30fc\u306bRNase A\u3092\u6dfb\u52a0\u3057\u3066\u3044\u308c\u3070\u3001\u554f\u984c\u306b\u306a\u308b\u3053\u3068\u306f\u3042\u308a\u307e\u305b\u3093\u3067\u3057\u305f\u3002<br \/>\n\u91cd\u8981\u306a\u70b9\u306fPCR\u53cd\u5fdc\u306b\u30c6\u30f3\u30d7\u30ec\u30fc\u30c8\u3092\u5165\u308c\u3059\u304e\u306a\u3044\u3053\u3068\u3067\u3059\u300220\u03bcl\u306e\u53cd\u5fdc\u7cfb\u306a\u30895\u304b\u308910ng\u3067\u5341\u5206\u3067\u3059\u3002\u5897\u5e45\u5b9f\u7e3e\u306e\u3042\u308b\u53cd\u5fdc\u7cfb\u3067\u3082\u300130ng\u4ee5\u4e0a\u306e\u30b2\u30ce\u30e0DNA\u3092\u5165\u308c\u308b\u3068\u4f55\u3082\u5897\u5e45\u3055\u308c\u306a\u304f\u306a\u308b\u3053\u3068\u3092\u3057\u3070\u3057\u3070\u7d4c\u9a13\u3057\u3066\u3044\u307e\u3059\u3002\u30d0\u30f3\u30c9\u304c\u307e\u3063\u305f\u304f\u73fe\u308c\u306a\u3044\u6642\u306f\u3001\u62bd\u51fa\u306e\u5931\u6557\u3067DNA\u304c\u5168\u304f\u5165\u3063\u3066\u3044\u306a\u3044\u304b\uff08\u62bd\u51fa\u3057\u305fDNA\u304c\u8cb4\u91cd\u3067\u96fb\u6c17\u6cf3\u52d5\u306b\u4f7f\u3046\u4f59\u88d5\u304c\u306a\u3044\u5834\u5408\uff09\u3001\u53cd\u5fdc\u306b\u4f7f\u3063\u305fDNA\u304c\u591a\u3059\u304e\u308b\u3068\u8003\u3048\u305f\u65b9\u304c\u3044\u3044\u3067\u3057\u3087\u3046\u3002\u3080\u3084\u307f\u306b\u30c6\u30f3\u30d7\u30ec\u30fc\u30c8\u3092\u5897\u3084\u3055\u306a\u3044\u3053\u3068\u304c\u809d\u8981\u3067\u3059\u3002<\/li>\n<li>GC\u30ea\u30c3\u30c1\u9818\u57df\u306e\u5897\u5e45\uff1a<br \/>\n\u5897\u5e45\u3055\u305b\u308b\u76ee\u7684\u306e\u914d\u5217\u306e\u4e2d\u306bGC\u30ea\u30c3\u30c1\u9818\u57df\u304c\u3042\u308b\u3068\u3001\u76f8\u88dc\u9396\u304c\u7d50\u5408\u3057\u3084\u3059\u304f\u30dd\u30ea\u30e1\u30e9\u30fc\u30bc\u304c\u8d70\u308a\u306b\u304f\u304f\u306a\u308b\u306e\u3067\u3001\u4f38\u5f35\u53cd\u5fdc\u306e\u904e\u7a0b\u3067\u3044\u304b\u306bDNA\u3092\u89e3\u96e2\u72b6\u614b\u306b\u4fdd\u3064\u304b\u304c\u30dd\u30a4\u30f3\u30c8\u3068\u306a\u308a\u307e\u3059\u3002\u305d\u306e\u305f\u3081\u3001\u53cd\u5fdc\u6e29\u5ea6\u3092\u9ad8\u304f\u3057\u3001\u9ad8\u3044\u53cd\u5fdc\u6e29\u5ea6\u3067\u3082\u5229\u7528\u3067\u304d\u308bTm\u5024\u306e\u9ad8\u3044\u30d7\u30e9\u30a4\u30de\u30fc\u3092\u8a2d\u8a08\u3059\u308b\u3053\u3068\u304c\u57fa\u672c\u3067\u3059\u3002GC\u30ea\u30c3\u30c1\u9818\u57df\u306b\u9650\u3089\u305a\u3001\u30b2\u30ce\u30e0PCR\u7528\u306e\u30d7\u30e9\u30a4\u30de\u30fc\u306eTm\u5024\u306f65\u2103\u4ee5\u4e0a\u306b\u8a2d\u8a08\u3059\u308b\u3053\u3068\u304c\u597d\u307e\u3057\u3044\u3067\u3059\u3002Tm\u5024\u304c65\u2103\u306e\u30d7\u30e9\u30a4\u30de\u30fc\u30bb\u30c3\u30c8\u3092\u7528\u3044\u308b\u5834\u5408\u3001\u6700\u521d\u306b60\u2103\u3067\u306e\u30a2\u30cb\u30fc\u30ea\u30f3\u30b0\u3092\u8a66\u3057\u307e\u3059\u3002\u3082\u3057\u3001\u76ee\u7684\u306e\u30d0\u30f3\u30c9\u3068\u3068\u3082\u306b\u30a8\u30af\u30b9\u30c8\u30e9\u30d0\u30f3\u30c9\u304c\u73fe\u308c\u305f\u308a\u3001\u30d0\u30f3\u30c9\u304c\u8584\u3044\u3068\u304d\u306f\u3001\u89e3\u6c7a\u7b56\u3068\u3057\u3066\u30a2\u30cb\u30fc\u30ea\u30f3\u30b0\u6e29\u5ea6\u3092\u4e0a\u3052\u308b\u3053\u3068\u3092\u8a66\u3057\u3066\u4e0b\u3055\u3044\u3002\u3053\u308c\u3067\u6539\u5584\u3055\u308c\u305f\u3053\u3068\u304c\u3042\u308a\u307e\u3057\u305f\u3002\u30a2\u30cb\u30fc\u30ea\u30f3\u30b0\u30b9\u30c6\u30c3\u30d7\u3092\u7701\u7565\u3057\u305f\uff12\u30b9\u30c6\u30c3\u30d7PCR\u30d7\u30ed\u30b0\u30e9\u30e0\u3082\u9078\u629e\u80a2\u3068\u3057\u3066\u8003\u616e\u3067\u304d\u307e\u3059\u3002<br \/>\n\u4e00\u65b9\u3001GC\u30ea\u30c3\u30c1\u9818\u57df\u306eTm\u5024\u3092\u4e0b\u3052\u308b\u305f\u3081\u3001\u53cd\u5fdc\u6db2\u306b5\u304b\u308910\uff05\u306eDMSO\u3092\u52a0\u3048\u308b\u624b\u6cd5\u3082\u52b9\u679c\u304c\u3042\u308a\u307e\u3057\u305f\u3002\u3053\u306e\u307b\u304b\u306b\u3001denature\uff08\u5909\u6027\uff09\u30b9\u30c6\u30c3\u30d7\u306e\u6e29\u5ea6\u3092\u901a\u5e38\u3088\u308a\uff12\u304b\u3089\uff13\u2103\u9ad8\u304f\u8a2d\u5b9a\u3059\u308b\u3053\u3068\u3067\u554f\u984c\u304c\u89e3\u6c7a\u3055\u308c\u305f\u3053\u3068\u3082\u7d4c\u9a13\u3057\u3066\u3044\u307e\u3059\u3002\u3053\u306e\u5834\u5408\u3001\u9175\u7d20\u306e\u5931\u6d3b\u3092\u9632\u3050\u305f\u3081denature\u30b9\u30c6\u30c3\u30d7\u306e\u6642\u9593\u3092\u77ed\u304f\u3057\u305f\u65b9\u304c\u7121\u96e3\u3067\u3059\u3002\u30b5\u30fc\u30de\u30eb\u30b5\u30a4\u30af\u30e9\u30fc\u306e\u6a5f\u7a2e\u306b\u3082\u4f9d\u308a\u307e\u3059\u304c\u3001\u6211\u3005\u306f20\u03bcL\u306e\u53cd\u5fdc\u7cfb\u306e\u5834\u540810\u79d2\u304f\u3089\u3044\u306b\u8a2d\u5b9a\u3057\u3066\u3044\u307e\u3059\u3002<\/li>\n<li>\u9577\u3044\u30b2\u30ce\u30e0\u9818\u57df\u306e\u5897\u5e45\uff1a<br \/>\n\u9577\u3044DNA\u9818\u57df\u3092\u30bf\u30fc\u30b2\u30c3\u30c8\u3068\u3057\u305f\u6642\u3001\u8981\u6c42\u3055\u308c\u308bPCR\u9175\u7d20\u306e\u5897\u5e45\u52b9\u7387\u3084\u71b1\u5b89\u5b9a\u6027\u304c\u683c\u6bb5\u306b\u9ad8\u304f\u306a\u308a\u307e\u3059\u3002\u660e\u78ba\u306a\u95be\u5024\u306f\u3042\u308a\u307e\u305b\u3093\u304c\u3001\u5897\u5e45\u9818\u57df\u304c10kb\u4ee5\u4e0a\u306e\u5834\u5408\u306fLong PCR\u5bfe\u5fdc\u3068\u8b33\u3063\u3066\u3044\u308bPCR\u8a66\u85ac\u3092\u9078\u3076\u3053\u3068\u3092\u304a\u52e7\u3081\u3057\u307e\u3059\u3002\u305f\u3060\u3057\u3001\u901a\u5e38\u306eDNA\u30dd\u30ea\u30e1\u30e9\u30fc\u30bc\u88fd\u54c1\u306e\u4e2d\u3067\u30826\u304b\u308910kb\u306e\u6bd4\u8f03\u7684\u9577\u3044\u30bf\u30fc\u30b2\u30c3\u30c8\u306e\u5897\u5e45\u306b\u9069\u7528\u3067\u304d\u308b\u3088\u3046\u306b\u3055\u308c\u305f\u3082\u306e\u304c\u3042\u308a\u307e\u3059\u3002\u6700\u65b0\u7248\u306ePCR\u9175\u7d20\u30ab\u30bf\u30ed\u30b0\u3092\u3088\u304f\u8aad\u3093\u3067\u9078\u3093\u3067\u304f\u3060\u3055\u3044\u3002\u3055\u3089\u306b\u3001\u300c\u88cf\u6280\u300d\u306e\u7bc4\u7587\u306b\u5165\u308a\u307e\u3059\u304c\u3001PCR\u7528\u9175\u7d20\u3068\u305d\u308c\u3068\u7570\u306a\u308b\u30d6\u30e9\u30f3\u30c9\u306e\u30d0\u30c3\u30d5\u30a1\u30fc\u3092\u4f75\u7528\u3059\u308b\u3053\u3068\u30678kb\u4ee5\u4e0a\u306e\u30d5\u30e9\u30b0\u30e1\u30f3\u30c8\u306e\u5897\u5e45\u306b\u6210\u529f\u3057\u305f\u3053\u3068\u3082\u3042\u308a\u307e\u3059\u3002<br \/>\nLong PCR\u3092\u884c\u3046\u6642\u306b\u9650\u3089\u305a\u3001\u53cd\u5fdc\u7cfb\u3084PCR\u30d7\u30ed\u30b0\u30e9\u30e0\u3092\u3044\u308d\u3044\u308d\u5909\u3048\u3066\u3082\u8907\u6570\u306e\u30d0\u30f3\u30c9\u307e\u305f\u306f\u30b9\u30e1\u30a2\u306a\u7523\u7269\u3057\u304b\u5f97\u3089\u308c\u306a\u3044\u3053\u3068\u3082\u3088\u304f\u3042\u308a\u307e\u3059\u3002\u3053\u306e\u5834\u5408\u3001\u4e00\u56de\u76ee\u306e\u53cd\u5fdc\u6db2\u30921\/50\u304b\u30891\/100\u306b\u5e0c\u91c8\u3057\u3001nested-PCR\u3092\u884c\u3046\u3053\u3068\u3067\u3001\u4e88\u60f3\u3088\u308a\u3082\u3046\u307e\u304f\u3044\u304f\u3053\u3068\u304c\u3042\u308a\u307e\u3057\u305f\u3002\u7570\u306a\u308b\u30d7\u30e9\u30a4\u30de\u30fc\u30bb\u30c3\u30c8\u3067nested-PCR\u3092\u884c\u3046\u306e\u3067\u306a\u304f\u3001first-PCR\u3068\u540c\u4e00\u306e\u30d7\u30e9\u30a4\u30de\u30fc\u30bb\u30c3\u30c8\u3092\u305d\u306e\u307e\u307e\u7528\u3044\u3066\u3082\u3001\u76ee\u7684\u30d5\u30e9\u30b0\u30e1\u30f3\u30c8\u306e\u307f\u304c\u5897\u5e45\u3055\u308c\u3066\u660e\u77ad\u306a\u5358\u4e00\u30d0\u30f3\u30c9\u304c\u898b\u3048\u3066\u304d\u305f\u3053\u3068\u3092\u4f55\u5ea6\u3082\u7d4c\u9a13\u3057\u3066\u3044\u307e\u3059\u3002(J.P.)<br \/>\n(Mail News 2007.03.27 \u63b2\u8f09\u8a18\u4e8b\u3088\u308a)<\/li>\n<\/ul>\n<p>&nbsp;<\/p>\n<h3>PCR\u306e\u8a71\u3057 (2)<\/h3>\n<ul style=\"line-height: 1.5em;\">\n<li>PCR \u3067\u6a19\u7684\u306e\u5897\u5e45\u304c\u898b\u3089\u308c\u306a\u3044\u3068\u304d\u306b\u3001\u92f3\u578bDNA\u6eb6\u6db2\u3092\u5e0c\u91c8\u3057\u3066\u4f7f\u7528\u3059\u308b\u3053\u3068\u3067\u5897\u5e45\u304c\u5f97\u3089\u308c\u308b\u3053\u3068\u304c\u3042\u308a\u307e\u3059\u3002\u30e9\u30dc\u30de\u30cb\u30e5\u30a2\u30eb\u300cPCR \u306e\u8a71\u3057 (1)\u300d\u3067\u89e6\u308c\u3066\u3044\u307e\u3059\u304c\u3001\u92f3\u578bDNA \u306e\u6eb6\u6db2\u4e2d\u306b\u5165\u308a\u8fbc\u3093\u3067\u3044\u308b\u4f55\u3089\u304b\u306e\u7269\u8cea\u304c\u3001PCR \u53cd\u5fdc\u3092\u963b\u5bb3\u3057\u3066\u3044\u308b\u3053\u3068\u304c\u8003\u3048\u3089\u308c\u3001\u6eb6\u6db2\u306e\u5e0c\u91c8\u306b\u3088\u308a\u305d\u306e\u963b\u5bb3\u7269\u8cea\u306e\u6fc3\u5ea6\u304c\u4e0b\u304c\u308b\u3053\u3068\u3067\u963b\u5bb3\u304c\u89e3\u9664\u3055\u308c\u308b\u3068\u8003\u3048\u3089\u308c\u307e\u3059\u3002<\/li>\n<li>\u3068\u3053\u308d\u3067\u3001\u305d\u306e\u963b\u5bb3\u7269\u8cea\u306f\u4f55\u3067\u3057\u3087\u3046\u304b\uff1f\u4eca\u56de\u3001\u300c<a href=\"https:\/\/brc.riken.jp\/dna\/GRP0005j\">BRC-JCM \u4fdd\u6709\u5fae\u751f\u7269\u682a\u7531\u6765\u30b2\u30ce\u30e0DNA<\/a>\u300d\u306e\u8abf\u88fd\u904e\u7a0b\u3067\u305d\u306e\u300c\u3053\u308c\u304c\u305d\u306e\u6b63\u4f53\u304b\uff1f\uff01\u300d\u3068\u3044\u3046\u4e8b\u4f8b\u306b\u906d\u9047\u3057\u307e\u3057\u305f\u306e\u3067\u3001\u5831\u544a\u3057\u307e\u3059\u3002<\/li>\n<li>\u907a\u4f1d\u5b50\u6750\u6599\u958b\u767a\u5ba4\u3067\u306f\u3001\u5fae\u751f\u7269\u6750\u6599\u958b\u767a\u5ba4\u3068\u5171\u540c\u306730 \u7a2e\u3042\u307e\u308a\u306e\u5fae\u751f\u7269DNA \u3092\u8abf\u88fd\u3057\u3001\u63d0\u4f9b\u3057\u3066\u304d\u307e\u3057\u305f\u3002\u5229\u7528\u8005\u304b\u3089\u65b0\u305f\u306a\u83cc\u306e\u8ffd\u52a0\u4f9d\u983c\u304c\u3042\u308a\u3001Mitsuokella multacida (JGD06653, JCM 2054T) \u3001Selenomonas ruminantium subsp. lactilytica (JGD06655, JCM 6582T) \u3068Prevotella albensis (JGD06677,JCM 12258T) \u306b\u3064\u3044\u3066\u3001\u3053\u308c\u307e\u3067\u3068\u540c\u3058\u3088\u3046\u306b\u30a2\u30eb\u30ab\u30ea\u6cd5\u3067DNA \u306e\u62bd\u51fa\u3092\u884c\u3044\u307e\u3057\u305f\u3002\uff08\u4e0b\u8a18\u53c2\u7167\uff09<\/li>\n<li>\u3044\u3064\u3082\u306e\u3088\u3046\u306b\u63d0\u4f9b\u7528\u306b\u8abf\u88fd\u3057\u305f\u30b5\u30f3\u30d7\u30eb\u309210\u500d\u5e0c\u91c8\u3057\u3001\u305d\u308c\u3092\u92f3\u578bDNA \u3068\u3057\u30661 uL \u3092\u7528\u3044\u305f20 uL\u306e\u53cd\u5fdc\u7cfb\u306716S rRNA gene \u3092\u6a19\u7684\u3068\u3059\u308b\u771f\u6b63\u7d30\u83cc\u691c\u67fb\u7528\u30d7\u30e9\u30a4\u30de\u30fc\u3092\u7528\u3044\u3001PCR \u3092\u884c\u3044\u307e\u3057\u305f\u304c\u3001\u5168\u304f\u30d0\u30f3\u30c9\u304c\u89b3\u5bdf\u3055\u308c\u307e\u305b\u3093\u3002\u3053\u308c\u307e\u3067\u8abf\u88fd\u3057\u3066\u304d\u305f30 \u3042\u307e\u308a\u306e\u682a\u3067\u306f\u3001\u3053\u306e\u3088\u3046\u306b\u6fc3\u3044\u92f3\u578bDNA \u3067PCR \u306b\u3088\u308b\u76ee\u7684\u30d0\u30f3\u30c9\u306e\u5897\u5e45\u963b\u5bb3\u304c\u304b\u304b\u308b\u83cc\u306f\u3042\u308a\u307e\u305b\u3093\u3067\u3057\u305f\u3002<\/li>\n<li>\u7591\u554f\u306b\u601d\u3044\u3001Mitsuokella multacida \u306eDNA \u62bd\u51fa\u904e\u7a0b\u3092\u601d\u3044\u51fa\u3057\u3066\u307f\u308b\u3068\u3001\u30a8\u30bf\u30ce\u30fc\u30eb\u6c88\u6bbf\u64cd\u4f5c\u306b\u3088\u3063\u3066\u3001DNA\u306e\u307b\u304b\u306b\u7d50\u6676\u69d8\u306e\u767d\u3044\u6c88\u6bbf\u304c\u89b3\u5bdf\u3055\u308c\u3066\u3044\u305f\u3053\u3068\u306b\u601d\u3044\u5f53\u305f\u308a\u307e\u3057\u305f\u3002\u3053\u306e\u767d\u3044\u7d50\u6676\u69d8\u306e\u6c88\u6bbf\u306f\u3001\u30a8\u30bf\u30ce\u30fc\u30eb\u6c88\u6bbf\u306e\u904e\u7a0b\u3067\u6790\u51fa\u3057\u3001\u6c34\u306b\u826f\u304f\u6eb6\u3051\u3001DNA \u3068\u6319\u52d5\u3092\u5171\u306b\u3057\u3066\u3044\u307e\u3059\u3002\u591a\u7cd6\u985e\u3067\u306f\u306a\u3044\u304b\u3068\u601d\u3044\u3001\u591a\u7cd6\u985e\u306e\u9664\u53bb\u306b\u6709\u52b9\u3067\u3042\u308b\u3068\u3055\u308c\u308bCTAB [Cety1 Trimethy1 AmmoniumBromide (hexadecyltrimethyl ammonium bromide)] \u306b\u3088\u308bDNA \u62bd\u51fa\u3092\u8a66\u307f\u307e\u3057\u305f\u3002\uff08\u4e0b\u8a18\u53c2\u7167\uff09<\/li>\n<li>\u305d\u306e\u5f8c\u3001\u901a\u5e38\u306e\u30a2\u30eb\u30ab\u30ea\u6cd5\u3067\u8abf\u88fd\u3057\u305fDNA \u3068\u305d\u308c\u3092\u3055\u3089\u306bCTAB \u62bd\u51fa\u3057\u3001\u591a\u7cd6\u985e\u3092\u9664\u304f\u64cd\u4f5c\u3092\u3057\u305fDNA \u3092\u540c\u3058\u6fc3\u5ea6 (300 ng\/uL) \u306b\u8abf\u88fd\u3057\u3001PCR \u306e\u92f3\u578b\u3068\u3057\u3066PCR \u3092\u884c\u3044\u307e\u3057\u305f\u3002\u4eca\u5ea6\u306f\u3001\u3082\u3068\u306eDNA \u309210 \u500d\u306b\u5e0c\u91c8\u3059\u308b\u3060\u3051\u3067\u306a\u304f\u300110 \u306e0.5 \u4e57\u500d\u305a\u3064\u6bb5\u968e\u5e0c\u91c8\u3057\u3001\u92f3\u578bDNA \u3068\u3057\u3066PCR \u3092\u884c\u3044\u307e\u3057\u305f\u3002PCR \u7523\u7269\u3092\u96fb\u6c17\u6cf3\u52d5\u306b\u3088\u308a\u78ba\u8a8d\u3057\u305f\u3068\u3053\u308d\u3001\u671f\u5f85\u306b\u53cd\u3057\u3066\u4e21\u8005\u3068\u3082\u540c\u3058\u7d50\u679c\u3068\u306a\u308a\u307e\u3057\u305f\u3002\u5373\u3061\u3001\u3082\u3068\u306eDNA \u309210 \u500d\u306b\u5e0c\u91c8\u3057\u305f30 ng\/uL \u306e\u92f3\u578bDNA \u30921 uL \u7528\u3044\u305f\u5834\u5408\u306f\u3001\u5897\u5e45\u3055\u308c\u308bDNA \u65ad\u7247\u306f\u89b3\u5bdf\u3055\u308c\u305a\u300110 \u306e1.5 \u4e57\u500d\u5e0c\u91c8\u3067\u306f\u8584\u3063\u3059\u3089\u3068\u3057\u305f\u30d0\u30f3\u30c9\u304c\u300110 \u306e2 \u4e57\u500d\u4ee5\u4e0a\u306e\u5e0c\u91c8\u3067\u826f\u597d\u306a\u30d0\u30f3\u30c9\u3092\u89b3\u5bdf\u3067\u304d\u307e\u3057\u305f\u3002<\/li>\n<li>\u7d50\u5c40\u3001CTAB \u62bd\u51fa\u306b\u3088\u308aDNA \u6eb6\u6db2\u304b\u3089\u7d50\u6676\u69d8\u306e\u767d\u3044\u6c88\u6bbf\u3092\u9664\u304f\u3053\u3068\u306f\u3067\u304d\u305f\u306e\u3067\u3059\u304c\u3001\u305d\u308c\u306b\u3088\u308aPCR\u306b\u3088\u308b\u5897\u5e45\u304c\u6539\u5584\u3055\u308c\u305f\u308f\u3051\u3067\u306f\u3042\u308a\u307e\u305b\u3093\u3067\u3057\u305f\u3002CTAB \u62bd\u51fa\u306b\u3088\u308a\u9664\u53bb\u3067\u304d\u305f\u3082\u306e\u304cPCR \u3092\u963b\u5bb3\u3057\u3066\u3044\u308b\u308f\u3051\u3067\u306f\u306a\u3044\u3053\u3068\u306f\u5206\u304b\u308a\u307e\u3057\u305f\u304c\u3001\u4f55\u304c\u963b\u5bb3\u7269\u8cea\u306a\u306e\u304b\u306f\u672a\u3060\u5206\u304b\u308a\u307e\u305b\u3093\u3002\u6539\u5584\u306e\u30d2\u30f3\u30c8\u3068\u306a\u308b\u60c5\u5831\u3092\u304a\u5f85\u3061\u3044\u305f\u3057\u3066\u304a\u308a\u307e\u3059\u3002(T.M.)<br \/>\n(Mail News 2008.10.10 \u63b2\u8f09\u8a18\u4e8b\u3088\u308a)<\/li>\n<\/ul>\n<h4><span style=\"text-decoration: underline;\">\u30a2\u30eb\u30ab\u30ea\u6cd5\u306b\u3088\u308b\u5fae\u751f\u7269\u30b2\u30ce\u30e0DNA \u62bd\u51fa<\/span><\/h4>\n<ol style=\"line-height: 1.5em;\">\n<li>50 mL \u30b3\u30cb\u30ab\u30eb\u30c1\u30e5\u30fc\u30d6\u3067\u83cc\u4f53\u30928 mL \u306e0.15 M NaCl + 0.1 M EDTA (pH8.0) \u306b\u61f8\u6fc1\u3059\u308b<\/li>\n<li>1 mL \u306e10 mg\/mL lysozyme (in PBS) \u3092\u52a0\u3048\u308b<\/li>\n<li>37\u2103\u6c34\u69fd\u30675 \u5206\u9593\u52a0\u6e29\u3059\u308b<\/li>\n<li>1 mL \u306e0.5 M Tris-HCl (pH7.5) + 5% (w\/v) SDS \u3092\u52a0\u3048\u308b<\/li>\n<li>25 uL \u306e10 mg\/mL proteinase K \u3092\u52a0\u3048\u308b<\/li>\n<li>60\u2103\u6c34\u69fd\u306710 \u5206\u9593\u52a0\u6e29\u3059\u308b<\/li>\n<li>\u7b49\u91cf\u306e\u30d5\u30a7\u30ce\u30fc\u30eb\u306b\u3088\u308b\u62bd\u51fa\u3092\uff12\u56de\u7e70\u308a\u8fd4\u3059<\/li>\n<li>\u7b49\u91cf\u306e\u30d5\u30a7\u30ce\u30fc\u30eb\uff0f\u30af\u30ed\u30ed\u30db\u30eb\u30e0\u306b\u3088\u308b\u62bd\u51fa\u3092\uff12\u56de\u7e70\u308a\u8fd4\u3059<\/li>\n<li>\u7b49\u91cf\u306e\u30af\u30ed\u30ed\u30db\u30eb\u30e0\u306b\u3088\u308b\u62bd\u51fa\u3092\uff11\u56de\u884c\u3046<\/li>\n<li>\u6c34\u76f8\u3092\u65b0\u3057\u3044\u30c1\u30e5\u30fc\u30d6\u306b\u79fb\u3057\u30012 \u500d\u91cf\u306e\u30a8\u30bf\u30ce\u30fc\u30eb\u3092\u52a0\u3048\u3001\u9759\u304b\u306b\u6df7\u548c\u3059\u308b<\/li>\n<li>\u73fe\u308c\u305fDNA \uff08\u767d\u3044\u7cf8\u304f\u305a\u306e\u584a\u72b6\u306e\u3082\u306e\uff09\u3092\u30d7\u30e9\u30b9\u30c1\u30c3\u30af\u88fd\u30a8\u30fc\u30bc\u306b\u5dfb\u304d\u53d6\u308a\u3001\u65b0\u3057\u3044\u30c1\u30e5\u30fc\u30d6\u306b\u79fb\u3059<\/li>\n<li>\u30a8\u30fc\u30bc\u304b\u3089DNA \u304c\u96e2\u308c\u306a\u3044\u5834\u5408\u306f\u3001\u7121\u7406\u306b\u3068\u3089\u305a\u306b\u30a8\u30fc\u30bc\u306e\u5148\u7aef\u3092\u30cf\u30b5\u30df\u3067\u5207\u308b<\/li>\n<li>10 mL \u306e75% \u30a8\u30bf\u30ce\u30fc\u30eb\u3092\u52a0\u3048\u3001\u6c88\u6bbf\u3092\u3059\u3059\u3050<\/li>\n<li>\u9060\u5fc3\u3092\u304b\u3051\u305a\u306b\u30a8\u30bf\u30ce\u30fc\u30eb\u3092\u5341\u5206\u9664\u304f<\/li>\n<li>DNA \u304c\u4e7e\u304b\u306a\u3044\u3046\u3061\u306b1 mL \u306eTE + 10 ug\/mL RNaseA \u3092\u52a0\u3048\u308b<\/li>\n<li>37\u2103\u306730 \u5206\u9593\u52a0\u6e29\u3059\u308b<\/li>\n<li>5 uL \u306e10 mg\/mL proteinase K \u3092\u52a0\u3048\u308b<\/li>\n<li>37\u2103\u30671 \u6642\u9593\u52a0\u6e29\u3059\u308b<\/li>\n<li>\u3055\u3089\u306b1 mL \u306eTE \u30d0\u30c3\u30d5\u30a1\u30fc\u3092\u52a0\u3048\u308b<\/li>\n<li>\u7b49\u91cf\u306e\u30d5\u30a7\u30ce\u30fc\u30eb\u306b\u3088\u308b\u62bd\u51fa\u3092\uff11\u56de\u884c\u3046<\/li>\n<li>\u7b49\u91cf\u306e\u30d5\u30a7\u30ce\u30fc\u30eb\uff0f\u30af\u30ed\u30ed\u30db\u30eb\u30e0\u306b\u3088\u308b\u62bd\u51fa\u3092\uff11\u56de\u884c\u3046<\/li>\n<li>\u7b49\u91cf\u306e\u30af\u30ed\u30ed\u30db\u30eb\u30e0\u306b\u3088\u308b\u62bd\u51fa\u3092\uff11\u56de\u884c\u3046<\/li>\n<li>\u6c34\u76f8\u3092\u65b0\u3057\u3044\u30c1\u30e5\u30fc\u30d6\u306b\u79fb\u3057\u30012 \u500d\u91cf\u306e\u30a8\u30bf\u30ce\u30fc\u30eb\u3092\u52a0\u3048\u3001\u9759\u304b\u306b\u6df7\u548c\u3059\u308b<\/li>\n<li>3,000rpm \u30675 \u5206\u9593\u9060\u5fc3\u5206\u96e2\u3092\u884c\u3046<\/li>\n<li>1 mL \u306e75% \u30a8\u30bf\u30ce\u30fc\u30eb\u3092\u52a0\u3048\u3001\u6c88\u6bbf\u3092\u3059\u3059\u3050<\/li>\n<li>3,000rpm \u30675 \u5206\u9593\u9060\u5fc3\u5206\u96e2\u3092\u884c\u3046<\/li>\n<li>\u30a8\u30bf\u30ce\u30fc\u30eb\u3092\u5341\u5206\u9664\u304f<\/li>\n<li>DNA \u304c\u4e7e\u304b\u306a\u3044\u3046\u3061\u306b1 mL \u306eTE \u30d0\u30c3\u30d5\u30a1\u30fc\u3092\u52a0\u3048\u3001\u5341\u5206\u6eb6\u89e3\u3055\u305b\u308b<\/li>\n<li>DNA \u6fc3\u5ea6\u3092\u6e2c\u5b9a\u3057\u3001\u6fc3\u3044\u5834\u5408\u306b\u306f\u3055\u3089\u306bTE \u30d0\u30c3\u30d5\u30a1\u30fc\u3092\u52a0\u3048\u308b<\/li>\n<\/ol>\n<h4><span style=\"text-decoration: underline;\">CTAB \u306b\u3088\u308bDNA \u62bd\u51fa<\/span><\/h4>\n<ol style=\"line-height: 1.5em;\">\n<li>TE \u30d0\u30c3\u30d5\u30a1\u30fc\u306b\u6eb6\u89e3\u3057\u305fDNA \u6eb6\u6db2600 uL \u3092\u65b0\u3057\u3044\u30c1\u30e5\u30fc\u30d6\u306b\u53d6\u5206\u3051\u308b<\/li>\n<li>100 uL \u306e5 M NaCl \u3092\u52a0\u3048\u308b<\/li>\n<li>Bacto-tryptone \u3000 \u3000 \u300012.0 g<\/li>\n<li>80 uL \u306eCTAB\/NaCl [10% (w\/v) CTAB\/0.7 M NaCl] \u6eb6\u6db2\u3092\u52a0\u3048\u308b<\/li>\n<li>65\u2103\u6c34\u69fd\u306710 \u5206\u9593\u52a0\u6e29\u3059\u308b<\/li>\n<li>\u7b49\u91cf\u306e\u30af\u30ed\u30ed\u30db\u30eb\u30e0\u306b\u3088\u308b\u62bd\u51fa\u3092\uff11\u56de\u884c\u3046<\/li>\n<li>\u7b49\u91cf\u306e\u30d5\u30a7\u30ce\u30fc\u30eb\uff0f\u30af\u30ed\u30ed\u30db\u30eb\u30e0\u306b\u3088\u308b\u62bd\u51fa\u3092\uff11\u56de\u884c\u3046<\/li>\n<li>\u7b49\u91cf\u306e\u30af\u30ed\u30ed\u30db\u30eb\u30e0\u306b\u3088\u308b\u62bd\u51fa\u3092\uff11\u56de\u884c\u3046<\/li>\n<li>\u6c34\u76f8\u3092\u65b0\u3057\u3044\u30c1\u30e5\u30fc\u30d6\u306b\u79fb\u3057\u30010.6 \u5bb9\u306e\u30a4\u30bd\u30d7\u30ed\u30d1\u30ce\u30fc\u30eb\u3092\u52a0\u3048\u3001\u9759\u304b\u306b\u6df7\u548c\u3059\u308b<\/li>\n<li>\u73fe\u308c\u305fDNA \uff08\u767d\u3044\u7cf8\u304f\u305a\u306e\u584a\u72b6\u306e\u3082\u306e\uff09\u3092\u30d7\u30e9\u30b9\u30c1\u30c3\u30af\u88fd\u30a8\u30fc\u30bc\u306b\u5dfb\u304d\u53d6\u308a\u3001\u65b0\u3057\u3044\u30c1\u30e5\u30fc\u30d6\u306b\u79fb\u3059<\/li>\n<li>\u30a8\u30fc\u30bc\u304b\u3089DNA \u304c\u96e2\u308c\u306a\u3044\u5834\u5408\u306f\u3001\u7121\u7406\u306b\u3068\u3089\u305a\u306b\u30a8\u30fc\u30bc\u306e\u5148\u7aef\u3092\u30cf\u30b5\u30df\u3067\u5207\u308b<\/li>\n<li>1 mL \u306e75% \u30a8\u30bf\u30ce\u30fc\u30eb\u3092\u52a0\u3048\u3001\u6c88\u6bbf\u3092\u3059\u3059\u3050<\/li>\n<li>3,000rpm \u30675 \u5206\u9593\u9060\u5fc3\u5206\u96e2\u3092\u884c\u3046<\/li>\n<li>\u30a8\u30bf\u30ce\u30fc\u30eb\u3092\u5341\u5206\u9664\u304f<\/li>\n<li>DNA \u304c\u4e7e\u304b\u306a\u3044\u3046\u3061\u306b300 uL \u306eTE \u30d0\u30c3\u30d5\u30a1\u30fc\u3092\u52a0\u3048\u3001\u5341\u5206\u6eb6\u89e3\u3055\u305b\u308b<\/li>\n<li>DNA \u6fc3\u5ea6\u3092\u6e2c\u5b9a\u3057\u3001\u6fc3\u3044\u5834\u5408\u306b\u306f\u3055\u3089\u306bTE \u30d0\u30c3\u30d5\u30a1\u30fc\u3092\u52a0\u3048\u308b<\/li>\n<\/ol>\n<p style=\"text-align:right;\">(MAN0012j 2008.10.10 T.M.)<\/p>\n<p style=\"text-align:left;\">2023.06.11<\/p>\n","protected":false},"excerpt":{"rendered":"<p>PCR\u306e\u8a71\u3057 (1) \u5e02\u8ca9\u306ePCR\u7528\u8a66\u85ac\u306e\u5897\u5e45\u52b9\u7387\u304c\u683c\u6bb5\u306b\u4e0a\u304c\u3063\u3066\u3044\u308b\u3068\u306f\u3044\u3048\u3001PCR\u3067\u76ee\u7684\u306e\u30d0\u30f3\u30c9\u304c\u73fe\u308c\u306a\u3044\u3053\u3068\u306f\u6642\u3005\u7d4c\u9a13\u3057\u307e\u3059\u3002\u30c6\u30f3\u30d7\u30ec\u30fc\u30c8\u304c\u30b2\u30ce\u30e0DNA\u3060\u3063\u305f\u308a\u3001\u5897\u5e45\u9818\u57df\u304c\u9577\u304b\u3063\u305f\u308a\u3001GC\u30ea\u30c3\u30c1\u306e\u914d\u5217\u304c\u542b\u307e\u308c\u308b\u5834\u5408\u306f\u6761\u4ef6\u8a2d\u5b9a\u304c\u7279\u306b\u96e3\u3057\u304f\u306a\u308a\u307e\u3059\u3002\u6700\u8fd1\u3001\u30b2\u30ce\u30df\u30c3\u30afPCR\u3092\u884c\u3046\u6a5f\u4f1a\u304c\u591a\u304f\u306a\u308a\u3001\u5931\u6557\u3068\u6210\u529f\u306e\u4f53\u9a13\u304b\u3089\u5e7e\u3064\u304b\u306e\u6559\u8a13\u3092\u5f97\u307e\u3057\u305f\u3002 \u30c6\u30f3\u30d7\u30ec\u30fc\u30c8\u306e\u6761\u4ef6\uff1a \u30b2\u30ce\u30e0DNA\u62bd\u51fa\u306b\u306f\u69d8\u3005\u306a\u30d7\u30ed\u30c8\u30b3\u30fc [&hellip;]<\/p>\n","protected":false},"author":9,"featured_media":0,"parent":296,"menu_order":0,"comment_status":"closed","ping_status":"closed","template":"","meta":{"_seopress_titles_title":"","_seopress_titles_desc":"","_seopress_robots_index":"","_seopress_robots_follow":"","_seopress_robots_imageindex":"","_seopress_robots_snippet":"","_seopress_robots_primary_cat":"","_seopress_robots_breadcrumbs":"","_seopress_robots_freeze_modified_date":"","_seopress_robots_custom_modified_date":"","_seopress_robots_canonical":"","_seopress_social_fb_title":"","_seopress_social_fb_desc":"","_seopress_social_fb_img":"","_seopress_social_fb_img_attachment_id":0,"_seopress_social_fb_img_width":0,"_seopress_social_fb_img_height":0,"_seopress_social_twitter_title":"","_seopress_social_twitter_desc":"","_seopress_social_twitter_img":"","_seopress_social_twitter_img_attachment_id":0,"_seopress_social_twitter_img_width":0,"_seopress_social_twitter_img_height":0,"_seopress_redirections_value":"","_seopress_redirections_enabled":"","_seopress_redirections_enabled_regex":"","_seopress_redirections_logged_status":"both","_seopress_redirections_param":"","_seopress_redirections_type":301,"_seopress_analysis_target_kw":"","footnotes":"","_wp_rev_ctl_limit":""},"class_list":["post-2418","page","type-page","status-publish","hentry"],"_links":{"self":[{"href":"http:\/\/dna.brc.riken.jp\/ja\/wp-json\/wp\/v2\/pages\/2418","targetHints":{"allow":["GET"]}}],"collection":[{"href":"http:\/\/dna.brc.riken.jp\/ja\/wp-json\/wp\/v2\/pages"}],"about":[{"href":"http:\/\/dna.brc.riken.jp\/ja\/wp-json\/wp\/v2\/types\/page"}],"author":[{"embeddable":true,"href":"http:\/\/dna.brc.riken.jp\/ja\/wp-json\/wp\/v2\/users\/9"}],"replies":[{"embeddable":true,"href":"http:\/\/dna.brc.riken.jp\/ja\/wp-json\/wp\/v2\/comments?post=2418"}],"version-history":[{"count":6,"href":"http:\/\/dna.brc.riken.jp\/ja\/wp-json\/wp\/v2\/pages\/2418\/revisions"}],"predecessor-version":[{"id":5642,"href":"http:\/\/dna.brc.riken.jp\/ja\/wp-json\/wp\/v2\/pages\/2418\/revisions\/5642"}],"up":[{"embeddable":true,"href":"http:\/\/dna.brc.riken.jp\/ja\/wp-json\/wp\/v2\/pages\/296"}],"wp:attachment":[{"href":"http:\/\/dna.brc.riken.jp\/ja\/wp-json\/wp\/v2\/media?parent=2418"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}