\u25a0\u3000\u5927\u8178\u83cc\u5f62\u8cea\u8ee2\u63db\u7528\u30b3\u30f3\u30d4\u30c6\u30f3\u30c8\u30bb\u30eb\u3092\u81ea\u4f5c\u3059\u308b\u5834\u5408\u306e\u30d7\u30ed\u30c8\u30b3\u30fc\u30eb\u3067\u3059\u3002\u7814\u7a76\u5ba4\u3084\u4eba\u306b\u3088\u3063\u3066\u4f5c\u308a\u65b9\u306f\u591a\u5c11\u9055\u3046\u3088\u3046\u3067\u3001\u3042\u308b\u7814\u7a76\u5ba4\u3067\u306f\u3059\u3079\u3066\u306e\u64cd\u4f5c\u3092\u4f4e\u6e29\u5ba4\u3067\u884c\u3046\u3088\u3046\u6559\u308f\u308a\u307e\u3057\u305f\u3002\u3053\u3053\u3067\u7d39\u4ecb\u3059\u308b\u306e\u306f\u3001\u79c1\u304cDNA Bank \u306b\u6765\u3066\u7fd2\u3063\u305f\u65b9\u6cd5\u3067\u3059\u3002\u3059\u3079\u3066\u306e\u64cd\u4f5c\u306f\u5ba4\u6e29\u3067\u884c\u3063\u3066\u69cb\u3044\u307e\u305b\u3093\u3002\u305f\u3060\u3057\u8a66\u85ac\u306f\u51b7\u3084\u3057\u305f\u3082\u306e\u3092\u4f7f\u3044\u3001\u9060\u5fc3\u6a5f\u306f4\u2103 \u306b\u8a2d\u5b9a\u3057\u3066\u304f\u3060\u3055\u3044\u3002\u52b9\u7387\u306f10^5\uff5e10^6 cfu \/ ug \u3068\u9ad8\u3044\u3082\u306e\u3068\u306f\u8a00\u3048\u307e\u305b\u3093\u304c\u3001\u77ed\u6642\u9593\u3067\u7c21\u5358\u306b\u4f5c\u308c\u308b\u65b9\u6cd5\u3067\u3059\u3002\u9ad8\u3044\u52b9\u7387\u304c\u5fc5\u8981\u306a\u30af\u30ed\u30fc\u30cb\u30f3\u30b0\u3084\u30e9\u30a4\u30d6\u30e9\u30ea\u30fc\u4f5c\u88fd\u306e\u7528\u9014\u306b\u306f\u5411\u3044\u3066\u3044\u307e\u305b\u3093\u304c\u3001\u7cbe\u88fd\u3057\u305f\u30d7\u30e9\u30b9\u30df\u30c9\u3092\u518d\u3073\u5897\u5e45\u3059\u308b\u5834\u5408\u3084\u30bf\u30f3\u30d1\u30af\u8cea\u767a\u73fe\u7528\u306e\u5f62\u8cea\u8ee2\u63db\u5927\u8178\u83cc\u306e\u6e96\u5099\u306b\u4f7f\u7528\u3059\u308b\u306a\u3089\u3001\u5168\u304f\u554f\u984c\u3042\u308a\u307e\u305b\u3093\u3002
\n\u25a0\u3000\u4ee5\u4e0b\u306b\u624b\u9806\u3092\u304a\u793a\u3057\u3057\u307e\u3059\u3002\u7528\u3044\u308b\u5668\u5177\u306f\u9069\u5b9c\u30aa\u30fc\u30c8\u30af\u30ec\u30fc\u30d6\u3092\u304b\u3051\u3001\u8a66\u85ac\u306f\u30aa\u30fc\u30c8\u30af\u30ec\u30fc\u30d6\u3001\u307e\u305f\u306f\u30d5\u30a3\u30eb\u30bf\u30fc\u306b\u3088\u308a\u6ec5\u83cc\u3057\u3066\u304a\u304d\u307e\u3059\u3002<\/p>\n
1\u65e5\u76ee\uff1a4mL \u306e\u57f9\u990a\u6db2\u306b\u690d\u83cc
\n2\u65e5\u76ee\uff1a\u4e0a\u3067\u6e96\u5099\u3057\u305f4 mL \u306e\u4e00\u6669\u57f9\u990a\u83cc\u3092400mL \u306e LB \u57f9\u5730\u306b\u690d\u3048\u308b
\n37\u2103\u3067OD600\u304c0.9\u304f\u3089\u3044\u307e\u3067\u57f9\u990a\uff081.0\u307e\u3067\u306f\u4e0a\u304c\u3089\u306a\u3044\u3088\u3046\u306b\uff09
\n200 mL \u00d72\u672c \u306b\u5206\u3051\u3066\u96c6\u83cc<\/p>\n
\uff1c\u4ee5\u4e0b\u306e\u64cd\u4f5c\u306f\u5206\u5272\u3057\u305f1\u672c\u305a\u3064\u306b\u3064\u3044\u3066\u8a18\u8ff0\u3057\u307e\u3059\uff1e<\/p>\n
1. 20mL \u306e 80 mM CaCl2, 50 mM MgCl2 \u3092\u52a0\u3048\u308b
\n2. \u30d4\u30da\u30c3\u30c6\u30a3\u30f3\u30b0\u3057\u305f\u61f8\u6fc1\u6db2\u309250 mL \u30c1\u30e5\u30fc\u30d6\u306b\u79fb\u3059
\n3. \u30dc\u30eb\u30c6\u30c3\u30af\u30b9\u3059\u308b\u300030\u79d2\uff5e1\u5206\u7a0b\u5ea6
\n4. \u6c37\u4e0a\u306b10\u5206\u653e\u7f6e
\n5. 3000 rpm, 4\u2103, 10\u5206\u3067\u96c6\u83cc<\/p>\n
1, 3, 4, 5\u306e\u64cd\u4f5c\u3092\u3082\u3046\u4e00\u5ea6\u884c\u3046<\/p>\n
\u96c6\u83cc\u3057\u305f\u6c88\u6bbf\u306b10mL \u306e 100mL CaCl2 \u3092\u52a0\u3048\u3001\u30dc\u30eb\u30c6\u30c3\u30af\u30b9
\n10mL \u306e 50% \u30b0\u30ea\u30bb\u30ed\u30fc\u30eb\u3092\u52a0\u3048\u3001\u30dc\u30eb\u30c6\u30c3\u30af\u30b9
\n\u30c1\u30e5\u30fc\u30d6\u306b\u9069\u91cf\u305a\u3064\u5206\u6ce8
\n\u76f4\u3061\u306b\u3001-80\u2103\u306b\u4fdd\u5b58<\/p>\n
\u25a0\u3000100\uff5e150 uL \u3092\u4e00\u3064\u306e\u30af\u30ed\u30fc\u30f3\u306b\u5bfe\u3057\u3066\u4f7f\u7528\u3057\u3066\u3044\u307e\u3059\u304c\u3001\u5206\u6ce8\u3057\u3066\u51b7\u51cd\u4fdd\u5b58\u3059\u308b\u91cf\u306f 0.5\uff5e1 mL \u3067\u3059\u3002\u4e00\u65e6\u7cbe\u88fd\u3057\u305f\u30d7\u30e9\u30b9\u30df\u30c9\u3092\u5897\u5e45\u3059\u308b\u306e\u3067\u3042\u308c\u3070\u3001\u30b3\u30f3\u30d4\u30c6\u30f3\u30c8\u30bb\u30eb\u3092\u6570\u56de\u51cd\u7d50\u878d\u89e3\u3057\u3066\u3082\u307e\u3060\u4f7f\u3048\u307e\u3059\u3002\u51cd\u7d50\u878d\u89e3\u306b\u3088\u308b\u52b9\u7387\u306e\u4f4e\u4e0b\u304c\u5fc3\u914d\u3067\u3042\u308c\u3070\u3001\u30b3\u30f3\u30d4\u30c6\u30f3\u30c8\u30bb\u30eb\u30921 mL \u3067\u51b7\u51cd\u3057\u3001\u521d\u56de\u4f7f\u7528\u6642\u306b\u5fc5\u8981\u91cf\u3092\u9664\u3044\u305f\u6b8b\u308a\u3092100 uL \u305a\u3064\u5206\u6ce8\u3057\u76f4\u3057\u3066\u51b7\u51cd\u3057\u3066\u304f\u3060\u3055\u3044\u3002
\n\u25a0\u3000\u30dc\u30eb\u30c3\u30c6\u30c3\u30af\u30b9\u3092\u304b\u3051\u3001\u6db2\u4f53\u7a92\u7d20\u3092\u4f7f\u7528\u3057\u306a\u3044\u3067\u4f5c\u88fd\u3059\u308b\u305f\u3081\u3001\u6700\u521d\u306f\u672c\u5f53\u306b\u3067\u304d\u308b\u306e\u304b\u4e0d\u5b89\u3067\u3057\u305f\u304c\u3001\u554f\u984c\u306a\u304f\u4f7f\u3063\u3066\u3044\u307e\u3059\u3002
\n\u25a0\u3000\u9ad8\u3044\u52b9\u7387\u306e\u30b3\u30f3\u30d4\u30c6\u30f3\u30c8\u30bb\u30eb\u3092\u5f97\u308b\u306e\u3067\u3042\u308c\u3070\u3001Inoue et al, 1990 \u3092\u304a\u52e7\u3081\u3057\u307e\u3059\u3002\u3053\u306e\u65b9\u6cd5\u3067\u306f\u3001\u83cc\u306e\u5897\u6b96\u3068\u8a66\u85ac\u306epH \u3092\u30b3\u30f3\u30c8\u30ed\u30fc\u30eb\u3067\u304d\u308c\u3070\u826f\u3044\u30b3\u30f3\u30d4\u30c6\u30f3\u30c8\u30bb\u30eb\u3092\u5f97\u308b\u3053\u3068\u304c\u3067\u304d\u307e\u3059\u3002Transformation buffer (TB) \u306e pH \u3092\u9069\u6b63\u306a\u3082\u306e\u306b\u5408\u308f\u305b\u308b\u306e\u304c\u96e3\u3057\u3044\u306e\u3067\u3059\u304c\u3001 pH \u306e\u7570\u306a\u308b\u4e8c\u3064\u306eTB \u3092\u8abf\u88fd\u3057\u3001\u305d\u308c\u3089\u306e\u6bd4\u7387\u3092\u5909\u3048\u3066\u6df7\u305c\u5408\u308f\u305b\u308b\u3053\u3068\u3067\uff08\u624b\u9593\u306f\u304b\u304b\u308a\u307e\u3059\u304c\uff09\u9069\u5207\u306apH \u306b\u5408\u3063\u305fTB \u3092\u5f97\u308b\u3053\u3068\u304c\u3067\u304d\u307e\u3059\u3002\u3053\u306e\u69d8\u306b pH \u306e\u7570\u306a\u308b2\u672c\u306e\u8a66\u85ac\u3092\u6e96\u5099\u3059\u308b\u65b9\u6cd5\u306f\u3001\u30ea\u30f3\u9178\u30ab\u30eb\u30b7\u30a6\u30e0\u6cd5\u306b\u3088\u308b\u52d5\u7269\u7d30\u80de\u3078\u306e\u907a\u4f1d\u5b50\u306e\u5c0e\u5165\u3067\u3082\u91cd\u5b9d\u3057\u307e\u3059\u3002(T.Y.)
\nInoue H, Nojima H, Okayama H. High efficiency transformation of Escherichia coli with plasmids. Gene 96 (1): 23-28, 1990. PMID: 2265755.<\/p>\n
(Mail News 2005.11.30 \u63b2\u8f09\u8a18\u4e8b\u3088\u308a)<\/p>\n","protected":false},"excerpt":{"rendered":"
\u5927\u8178\u83cc\u30b3\u30f3\u30d4\u30c6\u30f3\u30c8\u30bb\u30eb \u25a0\u3000\u5927\u8178\u83cc\u5f62\u8cea\u8ee2\u63db\u7528\u30b3\u30f3\u30d4\u30c6\u30f3\u30c8\u30bb\u30eb\u3092\u81ea\u4f5c\u3059\u308b\u5834\u5408\u306e\u30d7\u30ed\u30c8\u30b3\u30fc\u30eb\u3067\u3059\u3002\u7814\u7a76\u5ba4\u3084\u4eba\u306b\u3088\u3063\u3066\u4f5c\u308a\u65b9\u306f\u591a\u5c11\u9055\u3046\u3088\u3046\u3067\u3001\u3042\u308b\u7814\u7a76\u5ba4\u3067\u306f\u3059\u3079\u3066\u306e\u64cd\u4f5c\u3092\u4f4e\u6e29\u5ba4\u3067\u884c\u3046\u3088\u3046\u6559\u308f\u308a\u307e\u3057\u305f\u3002\u3053\u3053\u3067\u7d39\u4ecb\u3059\u308b\u306e\u306f\u3001\u79c1\u304cDNA Bank \u306b\u6765\u3066\u7fd2\u3063\u305f\u65b9\u6cd5\u3067\u3059\u3002\u3059\u3079\u3066\u306e\u64cd\u4f5c\u306f\u5ba4\u6e29\u3067\u884c\u3063\u3066\u69cb\u3044\u307e\u305b\u3093\u3002\u305f\u3060\u3057\u8a66\u85ac\u306f\u51b7\u3084\u3057\u305f\u3082\u306e\u3092\u4f7f\u3044\u3001\u9060\u5fc3\u6a5f\u306f4\u2103 \u306b\u8a2d\u5b9a\u3057\u3066\u304f\u3060\u3055\u3044\u3002\u52b9\u7387\u306f10^5\uff5e10^6 cfu \/ […]<\/p>\n","protected":false},"author":13,"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":"","_seopress_redirections_param":"","_seopress_redirections_type":0,"_seopress_analysis_target_kw":"","footnotes":"","_wp_rev_ctl_limit":""},"class_list":["post-4486","page","type-page","status-publish","hentry"],"_links":{"self":[{"href":"http:\/\/dna.brc.riken.jp\/ja\/wp-json\/wp\/v2\/pages\/4486","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\/13"}],"replies":[{"embeddable":true,"href":"http:\/\/dna.brc.riken.jp\/ja\/wp-json\/wp\/v2\/comments?post=4486"}],"version-history":[{"count":1,"href":"http:\/\/dna.brc.riken.jp\/ja\/wp-json\/wp\/v2\/pages\/4486\/revisions"}],"predecessor-version":[{"id":4487,"href":"http:\/\/dna.brc.riken.jp\/ja\/wp-json\/wp\/v2\/pages\/4486\/revisions\/4487"}],"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=4486"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}