{"id":520,"date":"2020-12-02T15:59:54","date_gmt":"2020-12-02T06:59:54","guid":{"rendered":"https:\/\/www.promega.co.jp\/resources\/faq\/?p=520"},"modified":"2021-02-15T17:58:01","modified_gmt":"2021-02-15T08:58:01","slug":"q187","status":"publish","type":"post","link":"https:\/\/www.promega.co.jp\/resources\/faq\/cloning\/enzyme\/restriction-enzyme\/re-enzyme-info\/q187","title":{"rendered":"\u5236\u9650\u9175\u7d20\u53cd\u5fdc\u3068\u306f?"},"content":{"rendered":"\n

\u5236\u9650\u9175\u7d20\u53cd\u5fdc\u306b\u3088\u308b\u89e3\u6790\u306b\u306f\u3001\u901a\u5e3820\u03bcl\u53cd\u5fdc\u6db2\u306b\u7d040.2\uff5e1.5\u03bcg \u306e\u57fa\u8ceaDNA\u306b\u5bfe\u3057\u30662\uff5e10\u500d\u306e\u904e\u5270\u91cf\u306e\u9175\u7d20(\u03bcg DNA\u3042\u305f\u308a2\uff5e10\u30e6\u30cb\u30c3\u30c8)\u3092\u52a0\u3048\u3066\u884c\u3044\u307e\u3059\u3002DNA\u3068\u9175\u7d20\u306e\u5bb9\u91cf\u306f\u3042\u308b\u30ec\u30d9\u30eb\u3092\u8d85\u3048\u306a\u3044\u7bc4\u56f2\u3067\u884c\u3063\u3066\u304f\u3060\u3055\u3044\u3002\u4f8b\u3048\u3070\u3001\u53cd\u5fdc\u6db2\u306e10%\u4ee5\u4e0a\u304cDNA\u30b5\u30f3\u30d7\u30eb\u306b\u306a\u3089\u306a\u3044\u3088\u3046\u306b\u3057\u3066\u304f\u3060\u3055\u3044\u3002DNA\u30b5\u30f3\u30d7\u30eb\u306b\u5236\u9650\u9175\u7d20\u53cd\u5fdc\u3092\u963b\u5bb3\u3059\u308b\u7269\u8cea(EDTA\u306a\u3069)\u304c\u542b\u307e\u308c\u3066\u3044\u308b\u53ef\u80fd\u6027\u304c\u3042\u308b\u305f\u3081\u3067\u3059\u3002\u305d\u306e\u305f\u3081\u3001\u30d7\u30ed\u30e1\u30ac\u3067\u306f20\u03bcl\u53cd\u5fdc\u6db2\u3042\u305f\u308a 2\u03bcl\u4ee5\u4e0b\u306eDNA\u3092\u4f7f\u3046\u3053\u3068\u3092\u63a8\u5968\u3057\u307e\u3059\u3002\u307e\u305f\u3001\u6700\u7d42\u7684\u306a\u30b0\u30ea\u30bb\u30ed\u30fc\u30eb\u6fc3\u5ea6\u306f5%\u4ee5\u4e0b\u304c\u671b\u307e\u3057\u3044\u306e\u3067\u3001\u9175\u7d20\u91cf\u306f\u53cd\u5fdc\u6db2\u306e10%\u307e\u305f\u306f\u305d\u308c\u4ee5\u4e0b\u306e\u91cf\u306b\u3059\u308b\u3053\u3068\u3092\u63a8\u5968\u3057\u307e\u3059\u3002\u30b0\u30ea\u30bb\u30ed\u30fc\u30eb\u6fc3\u5ea6\u304c\u9ad8\u3044\u5834\u5408\u3001\u9175\u7d20\u7fa4\u306b\u3088\u3063\u3066\u306fStar\u6d3b\u6027(\u901a\u5e38\u306e\u8a8d\u8b58\u90e8\u4f4d\u3068\u540c\u4e00\u3067\u306f\u306a\u304f\u985e\u4f3c\u3057\u305f\u914d\u5217\u3092\u6d88\u5316\u3059\u308b)\u3092\u751f\u3058\u308b\u3053\u3068\u304c\u3042\u308a\u307e\u3059\u3002<\/p>\n\n\n\n

\u4ee5\u4e0b\u306b\u5178\u578b\u7684\u306a\u5236\u9650\u9175\u7d20\u53cd\u5fdc\u306e\u4f8b\u3092\u793a\u3057\u307e\u3059\u3002\u5404\u8a66\u85ac\u306f\u4ee5\u4e0b\u306b\u793a\u3055\u308c\u305f\u9806\u3067\u6ec5\u83cc\u3057\u305f\u5fae\u91cf\u9060\u5fc3\u30c1\u30e5\u30fc\u30d6\u306b\u52a0\u3048\u3066\u304f\u3060\u3055\u3044\u3002<\/p>\n\n\n\n

\u6ec5\u83cc\u6e08\u307fNuclease-Free Water<\/td>14\u03bcl<\/td><\/tr>
\u9069\u5207\u306a\u5236\u9650\u9175\u7d20\u306e10X\u30d0\u30c3\u30d5\u30a1\u30fc(\u4f8b\u3048\u3070\u3001Buffer E)<\/td>2\u03bcl<\/td><\/tr>
\u30a2\u30bb\u30c1\u30eb\u5316BSA(1mg\/ml)*<\/td>2\u03bcl<\/td><\/tr>
DNA\u30b5\u30f3\u30d7\u30eb(0.2-1\u03bcg) in D.W.\u307e\u305f\u306fTE\u30d0\u30c3\u30d5\u30a1\u30fc<\/td>1\u03bcl<\/td><\/tr>
\u5236\u9650\u9175\u7d20(10u\/\u03bcl) (\u4f8b\u3048\u3070\u3001Bam<\/em>H I)<\/td>1\u03bcl<\/td><\/tr>
\u6700\u7d42\u5bb9\u91cf<\/td>20\u03bcl<\/td><\/tr><\/tbody><\/table><\/figure>\n\n\n\n

*\u30d7\u30ed\u30e1\u30ac\u306e\u5236\u9650\u9175\u7d20\u306b\u306f10mg\/ml\u306e\u30a2\u30bb\u30c1\u30eb\u5316BSA\u304c\u6dfb\u4ed8\u3055\u308c\u3066\u3044\u307e\u3059\u3002\u3053\u308c\u3092\u4f7f\u7528\u524d\u306bNuclease-Free Water\u306710\u500d\u5e0c\u91c8\u3057\u3066\u304f\u3060\u3055\u3044\u3002\u3082\u3057\u304f\u306f\u300110mg\/ml\u306e\u30a2\u30bb\u30c1\u30eb\u5316BSA(\u539f\u6db2)\u3092100\u500d\u5e0c\u91c8\u306b\u306a\u308b\u3088\u3046\u306b\u5236\u9650\u9175\u7d20\u53cd\u5fdc\u6db2\u306b\u76f4\u63a5\u52a0\u3048\u3066\u304f\u3060\u3055\u3044\u3002<\/p>\n\n\n\n

\u30d4\u30da\u30c3\u30c6\u30a3\u30f3\u30b0\u3067\u7a4f\u3084\u304b\u306b\u6df7\u305c(\u9175\u7d20\u3092\u52a0\u3048\u305f\u5f8c\u306b\u30dc\u30eb\u30c6\u30c3\u30af\u30b9\u306f\u884c\u308f\u306a\u3044\u3067\u304f\u3060\u3055\u3044)\u300112,000\u00d7g\u3067\u77ed\u6642\u9593\u306e\u9060\u5fc3\u3092\u884c\u3044\u3001\u6df7\u5408\u6db2\u3092\u30c1\u30e5\u30fc\u30d6\u306e\u5e95\u306b\u304a\u3068\u3057\u307e\u3059\u3002\u6700\u9069\u53cd\u5fdc\u6e29\u5ea6\u30671~4\u6642\u9593\u30a4\u30f3\u30ad\u30e5\u30d9\u30fc\u30c8\u3057\u307e\u3059(\u307b\u3068\u3093\u3069\u306e\u5236\u9650\u9175\u7d20\u306e\u6700\u9069\u53cd\u5fdc\u6e29\u5ea6\u306f37\u2103\u3067\u3059\u304c\u3001\u3042\u308b\u7a0b\u5ea6\u306e\u5e45\u3092\u6301\u305f\u305b\u308b\u3053\u3068\u304c\u3067\u304d\u307e\u3059)\u30024\u03bcl\u306eBlue\/Orange Loading Dye, 6X\u3092\u52a0\u3048\u3001\u96fb\u6c17\u6cf3\u52d5\u306b\u3088\u308a\u89e3\u6790\u3057\u307e\u3059\u3002<\/p>\n\n\n\n

\u5236\u9650\u9175\u7d20\u30d0\u30c3\u30d5\u30a1\u30fc\u306e\u7d44\u6210\u306b\u3064\u3044\u3066\u306f\u4e0b\u8a18pdf \u30d5\u30a1\u30a4\u30eb(Composition of Promega Restriction Enzyme Buffers [1X])\u3092\u3054\u89a7\u304f\u3060\u3055\u3044\u3002\u5404\u9175\u7d20\u3054\u3068\u53cd\u5fdc\u6e29\u5ea6\u3067\u306e4-CORE Buffer\u3092\u4f7f\u3063\u305f\u5236\u9650\u9175\u7d20\u306e\u76f8\u5bfe\u7684\u306a\u6d3b\u6027\u306b\u3064\u3044\u3066\u306f\u5236\u9650\u9175\u7d20\u95a2\u9023\u60c5\u5831\u306e\u8868\uff08page 18.16-17\uff09(Relative Activity of Restriction Enzymes in Promega’s 10X Buffers)\u3092\u3054\u89a7\u304f\u3060\u3055\u3044\u3002(eNotes-FAQ0001)<\/p>\n\n\n\n

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\u5236\u9650\u9175\u7d20\u95a2\u9023\u60c5\u5831\uff08pdf\u30d5\u30a1\u30a4\u30eb\uff09<\/a><\/p>\n","protected":false},"excerpt":{"rendered":"

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