{"id":55,"date":"2006-09-07T11:48:05","date_gmt":"2006-09-07T02:48:05","guid":{"rendered":"http:\/\/anatomy.med.gunma-u.ac.jp\/protocols\/?p=55"},"modified":"2015-02-25T21:31:47","modified_gmt":"2015-02-25T12:31:47","slug":"pcr","status":"publish","type":"post","link":"https:\/\/anatomy.med.gunma-u.ac.jp\/protocols\/?p=55","title":{"rendered":"PCR\u306b\u3088\u308b\u30a4\u30f3\u30b5\u30fc\u30c8\u306e\u30c1\u30a7\u30c3\u30af"},"content":{"rendered":"<h3>\u30b3\u30ed\u30cb\u30fc\u306e\u30d4\u30c3\u30af\u30a2\u30c3\u30d7<\/h3>\n<ol>\n<li>PCR\u30c1\u30e5\u30fc\u30d6\u306bLB\u309210 \u03bcl\u305a\u3064\u5206\u6ce8\u3059\u308b<\/li>\n<li>20 \u03bcl\u306e\u30de\u30a4\u30af\u30ed\u30d4\u30da\u30c3\u30c8\u306b\u6ec5\u83cc\u3057\u305f\u30c1\u30c3\u30d7\u3092\u53d6\u308a\u4ed8\u3051\u3001\u30c1\u30c3\u30d7\u306e\u5148\u7aef\u3092\u30b3\u30ed\u30cb\u30fc\u306b\u5f53\u3066\u3001\u659c\u3081\u4e0a\u306b\u3059\u304f\u3046\u3088\u3046\u306b\u30b3\u30ed\u30cb\u30fc\u3092\u62fe\u3046<\/li>\n<li>PCR\u30c1\u30e5\u30fc\u30d6\u306b\u30c1\u30c3\u30d7\u3092\u5dee\u3057\u8fbc\u307f\u3001LB\u3092\u6570\u56de\u30d4\u30da\u30c3\u30c6\u30a3\u30f3\u30b0\u3059\u308b<\/li>\n<li>4 \u2103\u3067\u4fdd\u5b58<\/li>\n<\/ol>\n<p><!--more--><\/p>\n<h3>\u30af\u30fc\u30eb\u30b9\u30bf\u30fc\u30c8PCR<\/h3>\n<ol>\n<li>\u975e\u7279\u7570\u7684\u53cd\u5fdc\u304c\u4f4e\u6e1b\u3055\u308c\u308b<\/li>\n<li>\u53cd\u5fdc\u6db2\u306e\u8abf\u88fd\u304b\u3089PCR\u53cd\u5fdc\u76f4\u524d\u307e\u3067\u6c37\u4e0a\u3067\u884c\u3046<\/li>\n<\/ol>\n<h3>PCR\u6df7\u5408\u6db2\u306e\u8abf\u88fd\uff0824\u691c\u4f53\u5206\uff09<\/h3>\n<ul>\n<li>167.4 \u03bcl H2O<\/li>\n<li>25 \u03bcl 10x buffer<\/li>\n<li>20 \u03bcl 2.5mM dNTPs<\/li>\n<li>1.3 \u03bcl Ex Taq<\/li>\n<li>11.3 \u03bcl 10 \u03bcM primer mix<\/li>\n<\/ul>\n<ul>\n<li>156.1 \u03bcl H2O<\/li>\n<li>25 \u03bcl 10x buffer<\/li>\n<li>20 \u03bcl 2.5mM dNTPs<\/li>\n<li>1.3 \u03bcl Ex Taq<\/li>\n<li>11.3 \u03bcl 10 \u03bcM right primer<\/li>\n<li>11.3 \u03bcl 10 \u03bcM left primer<\/li>\n<\/ul>\n<ul>\n<li>66.1 \u03bcl H2O<\/li>\n<li>25 \u03bcl 10x buffer<\/li>\n<li>20 \u03bcl 2.5mM dNTPs<\/li>\n<li>1.3 \u03bcl Ex Taq<\/li>\n<li>56.3 \u03bcl 2 \u03bcM right primer<\/li>\n<li>56.3 \u03bcl 2 \u03bcM left primer<\/li>\n<\/ul>\n<ul>\n<li>PCR\u30c1\u30e5\u30fc\u30d6\u306b9 \u03bcl\u305a\u3064\u5206\u6ce8\u3059\u308b<\/li>\n<\/ul>\n<h4>\u30d7\u30e9\u30a4\u30de\u30fc\u306e\u7d44\u307f\u5408\u308f\u305b\u4f8b<\/h4>\n<ul>\n<li>\u30d9\u30af\u30bf\u30fc\u306eMCS\u3092\u631f\u3080\u30d7\u30e9\u30a4\u30de\u30fc\u304b\u30a4\u30f3\u30b5\u30fc\u30c8\u306ePCR\u306b\u4f7f\u3063\u305f\u30d7\u30e9\u30a4\u30de\u30fc\u3092\u4f7f\u3046\n<ul>\n<li>T3 + T7\uff1apSC-B (StrataClone Blunt PCR Cloning Kit)\u3001pBlueScript<\/li>\n<li>SP6 + T7\uff1apCR-Blunt II-TOPO (Zero Blunt TOPO PCR Cloning Kit)<\/li>\n<li>HSP6 + CS2T7\uff1apCS2+ (\u30d7\u30e9\u30a4\u30de\u30fc\u914d\u5217\u304c\u7279\u6b8a)<\/li>\n<\/ul>\n<\/li>\n<li><span style=\"color: #ff0000;\">\u5c11\u306a\u304f\u3068\u3082\u4e00\u65b9\u306f\u30d9\u30af\u30bf\u30fc\u5074\u306e\u30d7\u30e9\u30a4\u30de\u30fc\u3092\u4f7f\u3046<\/span>\u3053\u3068\u3002\u30d5\u30ea\u30fc\u306e\u30a4\u30f3\u30b5\u30fc\u30c8\u304c\u5897\u5e45\u3055\u308c\u3066\u3057\u307e\u3046\u306e\u3092\u9632\u3050\u305f\u3081<\/li>\n<\/ul>\n<h3>PCR<\/h3>\n<ol>\n<li>2 \u03bcl\u306e\u30de\u30a4\u30af\u30ed\u30d4\u30da\u30c3\u30c8\u3092\u7528\u3044\u3066\u30b3\u30ed\u30cb\u30fc\u306e\u61f8\u6fc1\u6db2\u30921 \u03bcl\u3068\u308b<\/li>\n<li>\u3053\u308c\u3092PCR\u6df7\u5408\u6db2\u306e\u5165\u3063\u305fPCR\u30c1\u30e5\u30fc\u30d6\u306b\u52a0\u3048\u3001\u30c1\u30c3\u30d7\u306e\u5148\u3092\u6570\u56de\u307e\u308f\u3057\u3066\u6df7\u548c\u3059\u308b\uff08\u6ce1\u3092\u4f5c\u3089\u306a\u3044\u3088\u3046\u6ce8\u610f\u3002\u30c1\u30e5\u30fc\u30d6\u3092\u9060\u5fc3\u3057\u3066\u306f\u3044\u3051\u306a\u3044\uff09\n<ul>\n<li>\u30b3\u30c4\uff1a\u30de\u30a4\u30af\u30ed\u30d4\u30da\u30c3\u30c8\u30922\u6bb5\u968e\u5410\u304d\u51fa\u3059\u3068\u6ce1\u304c\u3067\u304d\u3066\u3057\u307e\u3046\u306e\u3067\u3001\u30b3\u30ed\u30cb\u30fc\u306e\u61f8\u6fc1\u6db2\u30921 \u03bcl\u3088\u308a\u591a\u3081\uff08\u301c1.2 \u03bcl\uff09\u306b\u3068\u308a\u30011\u6bb5\u968e\u3060\u3051\u5410\u304d\u51fa\u3059\u3068\u3088\u3044<\/li>\n<\/ul>\n<\/li>\n<\/ol>\n<h4>PCR\u30d7\u30ed\u30b0\u30e9\u30e0<\/h4>\n<ol>\n<li>4 \u2103, 30 s (\u30dd\u30fc\u30ba\u3057\u3066\u30c1\u30e5\u30fc\u30d6\u3092\u53d6\u308a\u4ed8\u3051\u308b\uff09<\/li>\n<li>98 \u2103, 2 min<\/li>\n<li>25\u30b5\u30a4\u30af\u30eb\n<ol>\n<li>98 \u2103, 10 s<\/li>\n<li>50 \u2103, 30 s<\/li>\n<li>72 \u2103, \u30a4\u30f3\u30b5\u30fc\u30c8\u9577 (kbp) \u00d7 1 min<\/li>\n<\/ol>\n<\/li>\n<li>72 \u2103, 7 min<\/li>\n<li>4 \u2103, \u221e<\/li>\n<\/ol>\n<h3>\u5224\u5b9a<\/h3>\n<ol>\n<li>6 \u00d7 Loading buffer\u30922 \u03bcl\u305a\u3064\u52a0\u3048\u306610 \u03bcl\u3092\u6cf3\u52d5\u3059\u308b<\/li>\n<li>\u30a4\u30f3\u30b5\u30fc\u30c8\u306e\u5927\u304d\u3055\u304b\u3089\u4e88\u6e2c\u3055\u308c\u308b\u5927\u304d\u3055\u306e\u30d0\u30f3\u30c9\u304c\u3067\u305f\u30b3\u30ed\u30cb\u30fc\u3092\u9078\u3073\u3001\u57f9\u990a\u3057\u3066\u4fdd\u5b58\u3059\u308b<\/li>\n<li>\u30a4\u30f3\u30b5\u30fc\u30c8\u306e\u65b9\u5411\u3092\u6c7a\u3081\u308b\u305f\u3081\u3001\u30a4\u30f3\u30b5\u30fc\u30c8\u3092\u5897\u5e45\u3057\u305f\u30d7\u30e9\u30a4\u30de\u30fc\u3068\u30d9\u30af\u30bf\u30fc\u306e\u30d7\u30e9\u30a4\u30de\u30fc\u3092\u305d\u308c\u305e\u308c\u4e00\u65b9\u305a\u30644\u901a\u308a\u7d44\u307f\u5408\u308f\u305b\u3066\u540c\u69d8\u306bPCR\u3092\u3059\u308b<\/li>\n<\/ol>\n<h3>\u5fc5\u8981\u306a\u3082\u306e<\/h3>\n<ul>\n<li>TaKaRa Ex Taq\uff08\u307e\u305f\u306f\u540c\u7b49\u54c1\uff09<\/li>\n<\/ul>\n<h3>\u3042\u308b\u3068\u3088\u3044\u3082\u306e<\/h3>\n<ul>\n<li><a href=\"http:\/\/www.fishersci.co.jp\/shop\/category\/category.aspx?category=20101026\">\u30d5\u30a3\u30c3\u30b7\u30e3\u30fc\u500b\u5225\u30ad\u30e3\u30c3\u30d7\u4ed88\u9023PCR\u30c1\u30e5\u30fc\u30d6<\/a>\uff08\u30ad\u30e3\u30c3\u30d7\u304c\u30aa\u30d5\u30bb\u30c3\u30c8\u3057\u3066\u308b\u306e\u3067\u3001\u30ad\u30e3\u30c3\u30d7\u3092\u958b\u3051\u305f\u30c1\u30e5\u30fc\u30d6\u3092\u67a0\u306b\u8a70\u3081\u3066\u306f\u3081\u3066\u3082\u3001\u6db2\u3092\u5165\u308c\u3089\u308c\u308b\uff09<\/li>\n<li><a href=\"http:\/\/anatomy.med.gunma-u.ac.jp\/protocols\/P2_001.pdf\">\u30a2\u30a4\u30b9\u30aa\u30f3<\/a>\uff08\u30af\u30fc\u30eb\u30b9\u30bf\u30fc\u30c8\u306b\u4fbf\u5229\uff09<\/li>\n<\/ul>\n","protected":false},"excerpt":{"rendered":"<p>\u30b3\u30ed\u30cb\u30fc\u306e\u30d4\u30c3\u30af\u30a2\u30c3\u30d7 PCR\u30c1\u30e5\u30fc\u30d6\u306bLB\u309210 \u03bcl\u305a\u3064\u5206\u6ce8\u3059\u308b 20 \u03bcl\u306e\u30de\u30a4\u30af\u30ed\u30d4\u30da\u30c3\u30c8\u306b\u6ec5\u83cc\u3057\u305f\u30c1\u30c3\u30d7\u3092\u53d6\u308a\u4ed8\u3051\u3001\u30c1\u30c3\u30d7\u306e\u5148\u7aef\u3092\u30b3\u30ed\u30cb\u30fc\u306b\u5f53\u3066\u3001\u659c\u3081\u4e0a\u306b\u3059\u304f\u3046\u3088\u3046\u306b\u30b3\u30ed\u30cb\u30fc\u3092\u62fe\u3046 PCR\u30c1\u30e5\u30fc\u30d6\u306b\u30c1\u30c3\u30d7\u3092\u5dee\u3057<\/p>\n","protected":false},"author":3,"featured_media":0,"comment_status":"open","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"inline_featured_image":false,"_jetpack_newsletter_access":"","_jetpack_dont_email_post_to_subs":false,"_jetpack_newsletter_tier_id":0,"_jetpack_memberships_contains_paywalled_content":false,"_jetpack_memberships_contains_paid_content":false,"footnotes":"","jetpack_publicize_message":"","jetpack_publicize_feature_enabled":true,"jetpack_social_post_already_shared":false,"jetpack_social_options":{"image_generator_settings":{"template":"highway","default_image_id":0,"font":"","enabled":false},"version":2},"jetpack_post_was_ever_published":false},"categories":[4],"tags":[],"class_list":["post-55","post","type-post","status-publish","format-standard","hentry","category-molecular-biology"],"jetpack_publicize_connections":[],"jetpack_featured_media_url":"","jetpack_sharing_enabled":true,"jetpack_shortlink":"https:\/\/wp.me\/s4fFyn-pcr","jetpack-related-posts":[],"_links":{"self":[{"href":"https:\/\/anatomy.med.gunma-u.ac.jp\/protocols\/index.php?rest_route=\/wp\/v2\/posts\/55","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/anatomy.med.gunma-u.ac.jp\/protocols\/index.php?rest_route=\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/anatomy.med.gunma-u.ac.jp\/protocols\/index.php?rest_route=\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/anatomy.med.gunma-u.ac.jp\/protocols\/index.php?rest_route=\/wp\/v2\/users\/3"}],"replies":[{"embeddable":true,"href":"https:\/\/anatomy.med.gunma-u.ac.jp\/protocols\/index.php?rest_route=%2Fwp%2Fv2%2Fcomments&post=55"}],"version-history":[{"count":4,"href":"https:\/\/anatomy.med.gunma-u.ac.jp\/protocols\/index.php?rest_route=\/wp\/v2\/posts\/55\/revisions"}],"predecessor-version":[{"id":400,"href":"https:\/\/anatomy.med.gunma-u.ac.jp\/protocols\/index.php?rest_route=\/wp\/v2\/posts\/55\/revisions\/400"}],"wp:attachment":[{"href":"https:\/\/anatomy.med.gunma-u.ac.jp\/protocols\/index.php?rest_route=%2Fwp%2Fv2%2Fmedia&parent=55"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/anatomy.med.gunma-u.ac.jp\/protocols\/index.php?rest_route=%2Fwp%2Fv2%2Fcategories&post=55"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/anatomy.med.gunma-u.ac.jp\/protocols\/index.php?rest_route=%2Fwp%2Fv2%2Ftags&post=55"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}