{"id":83,"date":"2015-09-18T00:00:00","date_gmt":"2015-09-18T00:00:00","guid":{"rendered":"https:\/\/www.thermofisher.com\/blog\/learning-at-the-bench\/checking-your-t4-dna-ligase-activity\/"},"modified":"2024-08-27T11:43:41","modified_gmt":"2024-08-27T02:43:41","slug":"checking-your-t4-dna-ligase-activity","status":"publish","type":"post","link":"https:\/\/www.thermofisher.com\/blog\/learning-at-the-bench\/checking-your-t4-dna-ligase-activity\/","title":{"rendered":"\u30e9\u30a4\u30b2\u30fc\u30b7\u30e7\u30f3\u304c\u5931\u6557\u3057\u305f\u3068\u304d\u306b\u8a66\u3057\u305f\u3044\uff01\u30ea\u30ac\u30fc\u30bc\u306e\u6d3b\u6027\u3092\u78ba\u8a8d\u3059\u308b\u65b9\u6cd5"},"content":{"rendered":"

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\u6982\u8981<\/h2>\n

SDS\u542b\u6709\u306e\u30ed\u30fc\u30c7\u30a3\u30f3\u30b0\u30c0\u30a4(SDS\u306e\u6700\u7d42\u6fc3\u5ea6\u306f0.2%)\u3068\u6df7\u5408\u3057\u3001\u901a\u5e38\u306e\u30a2\u30ac\u30ed\u30fc\u30b9\u30b2\u30eb\u96fb\u6c17\u6cf3\u52d5\u306b\u6d41\u3059\u3060\u3051\u3067\u3001\u30e9\u30a4\u30b2\u30fc\u30b7\u30e7\u30f3\u53cd\u5fdc\u306e\u52b9\u7387\u3092\u78ba\u8a8d\u3067\u304d\u307e\u3059\u3002\u30e9\u30a4\u30b2\u30fc\u30b7\u30e7\u30f3\u53cd\u5fdc\u5f8c\u306b\u3044\u304f\u3064\u304b\u306e\u9ad8\u5206\u5b50\u306e\u30d0\u30f3\u30c9\u304c\u73fe\u308c\u308b\u306f\u305a\u3067\u3059\u3002\u30ed\u30fc\u30c7\u30a3\u30f3\u30b0\u30c0\u30a4\u306bSDS\u304c\u542b\u307e\u308c\u3066\u3044\u306a\u3044\u3068\u3001\u30b9\u30e1\u30a2\u30d0\u30f3\u30c9\u304c\u51fa\u73fe\u3057\u3001\u30ea\u30ac\u30fc\u30bc\u306e\u6d3b\u6027\u3092\u6e2c\u308b\u3053\u3068\u304c\u96e3\u3057\u304f\u306a\u308a\u307e\u3059\u3002\uff08\u56f31\uff09<\/p>\n

\"Checking-Your-T4-DNA-Ligase-Activity-fig1\"<\/a><\/p>\n

\u56f31
\n\u30ec\u30fc\u30f31\uff1a\u30e9\u30a4\u30b2\u30fc\u30b7\u30e7\u30f3\u53cd\u5fdc\u524d\u306e\u30b5\u30f3\u30d7\u30eb
\n\u30ec\u30fc\u30f32\uff1a\u30e9\u30a4\u30b2\u30fc\u30b7\u30e7\u30f3\u53cd\u5fdc\u5f8c\u3001SDS\u4e0d\u542b\u6709\u306e\u30ed\u30fc\u30c7\u30a3\u30f3\u30b0\u30c0\u30a4\u3092\u4f7f\u7528\u3057\u305f\u3082\u306e
\n\u30ec\u30fc\u30f33\uff1a\u30e9\u30a4\u30b2\u30fc\u30b7\u30e7\u30f3\u53cd\u5fdc\u5f8c\u3001SDS\u542b\u6709\u306e\u30ed\u30fc\u30c7\u30a3\u30f3\u30b0\u30c0\u30a4\u3092\u4f7f\u7528\u3057\u305f\u3082\u306e
\n\u30ec\u30fc\u30f34\uff1aGeneRuler DNA\u30e9\u30c0\u30fc\u30df\u30c3\u30af\u30b9<\/a><\/p>\n

 <\/p>\n

\u57fa\u8ceaDNA\u3068\u3057\u3066\u3001HindIII\u3067\u5207\u65ad\u3055\u308c\u305f\u30e9\u30e0\u30c0DNA<\/a>\u3092\u4f7f\u7528\u3059\u308b\u3053\u3068\u3067\u3082\u30ea\u30ac\u30fc\u30bc\u306e\u6d3b\u6027\u3092\u6e2c\u308b\u3053\u3068\u304c\u3067\u304d\u307e\u3059\u3002\u3053\u306eDNA\u306f\u9ad8\u7d14\u5ea6\u306e\u305f\u3081\u3001\u4e0b\u8a18\u306e\u30d7\u30ed\u30c8\u30b3\u30eb\u3092\u884c\u3046\u306e\u304c\u7c21\u5358\u306b\u306a\u308a\u307e\u3059\u3002<\/p>\n

\u30d7\u30ed\u30c8\u30b3\u30eb<\/h2>\n

0.5 \u03bcg HindIII\u3067\u5207\u65ad\u3055\u308c\u305f\u30e9\u30e0\u30c0DNA<\/a>\u3068
\n2 \u03bcL 10x T4 DNA Ligase buffer<\/a>\u3068
\n1\u30e6\u30cb\u30c3\u30c8\u306eT4 DNA\u30ea\u30ac\u30fc\u30bc<\/a>\u3092\u8ffd\u52a0\u3057\u307e\u3059 (1\u00d7T4 DNA Ligase buffer\u3067\u5e0c\u91c8\u304c\u53ef\u80fd\u3067\u3059\uff09
\n\u5168\u6eb6\u6db2\u91cf\u304c20 \u03bcL\u306b\u306a\u308b\u3088\u3046\u306b\u6c34\u3067\u8abf\u88fd\u3057\u300122\u00b0C (\u5ba4\u6e29)\u306710\u5206\u9593\u53cd\u5fdc\u3055\u305b\u307e\u3059\u3002<\/p>\n

\u7d50\u679c<\/h2>\n

\u4e0a\u8a18\u3067\u8ff0\u3079\u305f\u3088\u3046\u306b\u3001SDS\u542b\u6709\u306e\u30ed\u30fc\u30c7\u30a3\u30f3\u30b0\u30c0\u30a4\u3092\u4f7f\u7528\u3057\u3066\u300110 \u03bcL\u306e\u53cd\u5fdc\u6db2\u3092\u30a2\u30ac\u30ed\u30fc\u30b9\u30b2\u30eb\u96fb\u6c17\u6cf3\u52d5\u306b\u6d41\u3057\u307e\u3059\u3002\u30e9\u30a4\u30b2\u30fc\u30b7\u30e7\u30f3\u53cd\u5fdc\u5f8c\u306e\u30b5\u30f3\u30d7\u30eb\u3067\u306f\u3001\u56f3 2\u3067\u793a\u3059\u3088\u3046\u306b\u3044\u304f\u3064\u304b\u306e\u9ad8\u5206\u5b50\u306e\u30d0\u30f3\u30c9\u304c\u73fe\u308c\u308b\u306f\u305a\u3067\u3059\u3002<\/p>\n

\"Checking-Your-T4-DNA-Ligase-Activity-fig2\"<\/a>\u56f32
\n\u30ec\u30fc\u30f31\uff1a\u30e9\u30a4\u30b2\u30fc\u30b7\u30e7\u30f3\u53cd\u5fdc\u524d\u306e\u30e9\u30e0\u30c0DNA
\n\u30ec\u30fc\u30f32\uff1a\u30e9\u30a4\u30b2\u30fc\u30b7\u30e7\u30f3\u53cd\u5fdc\u5f8c\u306e\u30e9\u30e0\u30c0DNA<\/p>\n

 <\/p>\n

 <\/p>\n

 <\/p>\n

\u30e9\u30e0\u30c0DNA\u3092\u4f7f\u7528\u3057\u305f\u5834\u5408\u306b\u306f\u30e9\u30a4\u30b2\u30fc\u30b7\u30e7\u30f3\u306f\u3046\u307e\u304f\u3044\u304f\u304c\u3001\u76ee\u7684\u306e\u30b5\u30f3\u30d7\u30eb\u3092\u4f7f\u7528\u3057\u305f\u969b\u306b\u30e9\u30a4\u30b2\u30fc\u30b7\u30e7\u30f3\u304c\u3046\u307e\u304f\u3044\u304b\u306a\u3044\u5834\u5408\u306f\u3001\u30ea\u30ac\u30fc\u30bc\u306b\u306f\u554f\u984c\u306f\u306a\u304f\u3001\u30b5\u30f3\u30d7\u30eb\u306b\u554f\u984c\u304c\u3042\u308b\u3068\u8003\u3048\u3089\u308c\u307e\u3059\u3002\u3053\u308c\u306b\u306f\u3001\u3044\u304f\u3064\u304b\u306e\u7406\u7531\u304c\u539f\u56e0\u3067\u3042\u308b\u53ef\u80fd\u6027\u304c\u3042\u308a\u307e\u3059\u3002\u5f0a\u793e\u306e\u30c6\u30af\u30cb\u30ab\u30eb\u30b5\u30dd\u30fc\u30c8<\/a>\u306f\u3001\u3053\u306e\u3088\u3046\u306a\u30c8\u30e9\u30d6\u30eb\u30b7\u30e5\u30fc\u30c6\u30a3\u30f3\u30b0\u3092\u652f\u63f4\u3044\u305f\u3057\u307e\u3059\u3002<\/p>\n

\u3053\u308c\u3060\u3051\u306f\u77e5\u3063\u3066\u304a\u304d\u305f\u3044\u30af\u30ed\u30fc\u30cb\u30f3\u30b0\u306e\u57fa\u790e\u77e5\u8b58\u3001\u30b3\u30b3\u306b\u3042\u308a\u307e\u3059\uff01<\/h2>\n

Invitrogen \u5206\u5b50\u751f\u7269\u5b66\u6559\u5ba4\u3067\u306f\u3001\u5206\u5b50\u751f\u7269\u5b66\u306e\u6559\u80b2\u306e\u30e1\u30a4\u30f3\u3068\u3057\u3066\u3001\u521d\u5fc3\u8005\u3060\u3051\u3067\u306a\u304f\u7d4c\u9a13\u306e\u3042\u308b\u5206\u5b50\u751f\u7269\u5b66\u8005\u306e\u305f\u3081\u306b\u3001\u8c4a\u5bcc\u3067\u4fe1\u983c\u3067\u304d\u308b\u6280\u8853\u7684\u306a\u5185\u5bb9\u3092\u3054\u63d0\u4f9b\u3057\u3066\u3044\u307e\u3059\u3002\u57fa\u790e\u77e5\u8b58\u3092\u5b66\u3076\u6642\u306b\u306f\u3044\u3064\u3067\u3082\u3001Invitrogen \u5206\u5b50\u751f\u7269\u5b66\u6559\u5ba4\u306e\u7121\u6599PDF\u3092\u6d3b\u7528\u3057\u3066\u304f\u3060\u3055\u3044\uff01<\/p>\n

\u30c0\u30a6\u30f3\u30ed\u30fc\u30c9\u3059\u308b<\/a><\/p>\n

 <\/p>\n

 <\/p>\n

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