{"id":5374,"date":"2022-04-15T10:38:04","date_gmt":"2022-04-15T01:38:04","guid":{"rendered":"https:\/\/gikenbio.com\/2025test\/?page_id=5374"},"modified":"2025-01-21T17:37:36","modified_gmt":"2025-01-21T08:37:36","slug":"mifish","status":"publish","type":"page","link":"https:\/\/gikenbio.com\/2025test\/dnaanalysis\/ngs\/amplicon\/techinfo\/mifish\/","title":{"rendered":"12S rRNA (cMiFish) \u30d7\u30e9\u30a4\u30de\u30fc\u60c5\u5831"},"content":{"rendered":"<p>\u25cf\u30d7\u30e9\u30a4\u30de\u30fc\u914d\u5217\uff085\u2019\u21923\u2019\uff09\/\u30a2\u30f3\u30d7\u30ea\u30b3\u30f3\u30b7\u30fc\u30b1\u30f3\u30b9\u89e3\u6790\uff08illumina MiSeq\uff09\u7528<br \/>\n1st-cMiFish F\uff1a<br \/>\n\u2460ACACTCTTTCCCTACACGACGCTCTTCCGATCT-<span style=\"color: #ff0000;\">NNNNNN<\/span>&#8211;<strong>RGTTGGTAAATCTCGTGCCAGC<\/strong><br \/>\n\u2461ACACTCTTTCCCTACACGACGCTCTTCCGATCT-<span style=\"color: #ff0000;\">NNNNNN<\/span>&#8211;<strong>GTCGGTAAAACTCGTGCCAGC<\/strong><br \/>\n\u2462ACACTCTTTCCCTACACGACGCTCTTCCGATCT-<span style=\"color: #ff0000;\">NNNNNN<\/span>&#8211;<strong>GCCGGTAAAACTCGTGCCAGC<\/strong><br \/>\n\u2463ACACTCTTTCCCTACACGACGCTCTTCCGATCT-<strong>GCCGGTTAATCTCGTGCCAGC<\/strong><br \/>\n1st-cMiFish R\uff1a<br \/>\n\u2460GTGACTGGAGTTCAGACGTGTGCTCTTCCGATCT-<span style=\"color: #ff0000;\">NNNNNN<\/span>&#8211;<strong>GCATAGTGGGGTATCTAATCCTAGTTTG<br \/>\n<\/strong>\u2461GTGACTGGAGTTCAGACGTGTGCTCTTCCGATCT-<span style=\"color: #ff0000;\">NNNNNN<\/span>&#8211;<strong>CATAGTGGGGTATCTAATCCCAGTTTG<br \/>\n<\/strong>\u2462GTGACTGGAGTTCAGACGTGTGCTCTTCCGATCT-<span style=\"color: #ff0000;\">NNNNNN<\/span>&#8211;<strong>CATAGGAGGGTGTCTAATCCCCGTTTG<br \/>\n<\/strong>\u2463GTGACTGGAGTTCAGACGTGTGCTCTTCCGATCT-<strong>CATAGTGGGGTATCTAATCCCAGTTTG<br \/>\n<\/strong>\u203b\u305d\u308c\u305e\u308c\u2460\uff1a\u2461\uff1a\u2462\uff1a\u2463=2\uff1a4\uff1a2\uff1a1\u3067\u6df7\u5408\u3057\u305f\u30d7\u30e9\u30a4\u30de\u30fc\u3067\u3059\u3002<strong><br \/>\n<\/strong><\/p>\n<p class=\"well\">\u7d30\u5b57\uff1a\u30a2\u30c0\u30d7\u30bf\u30fc\u914d\u5217\uff082nd PCR\u7528\u306e\u30d7\u30e9\u30a4\u30de\u30fc\u3068\u306e\u76f8\u540c\u90e8\u5206\uff09<br \/>\n<span style=\"color: #ff0000;\">\u8d64\u5b57 N<\/span>\uff1a\u30b7\u30fc\u30b1\u30f3\u30b9\u89e3\u6790\u6642\u306b\u304a\u3051\u308b\u54c1\u8cea\u5411\u4e0a\u3092\u76ee\u7684\u3068\u3057\u3066\u30011-6\u5869\u57fa\u306e\u7570\u306a\u308b\u9577\u3055\u306e\u30e9\u30f3\u30c0\u30e0\u914d\u5217\u304c\u633f\u5165\u3055\u308c\u305f\u6df7\u5408\u30d7\u30e9\u30a4\u30de\u30fc\u3067\u3059<br \/>\n<strong>\u592a\u5b57<\/strong>\uff1a12S rRNA \u3068\u306e\u76f8\u540c\u914d\u5217<\/p>\n<p>\u25cf\u53c2\u8003\u6587\u732e<br \/>\nMiya M et al. R. Soc. open sci. (2015) 2:150088<\/p>\n<p>\u25cf1stPCR\u5f8c\u306e\u5897\u5e45\u9577<br \/>\n\u7d04310 bp<\/p>\n<h5 class=\"style5a\">1stPCR\u306e\u53cd\u5fdc\u7d44\u6210\u3068\u53cd\u5fdc\u6761\u4ef6\u4f8b<\/h5>\n<p>\u4ee5\u4e0b\u306f\u3001 PCR \u9175\u7d20\u306b ExTaq HS [TaKaRa]\u3092\u4f7f\u7528\u3057\u305f\u4f8b\u3067\u3059\u3002<br \/>\n\u4ed6\u306e PCR \u9175\u7d20\u3092\u4f7f\u7528\u3059\u308b\u5834\u5408\u306f\u3001\u305d\u306ePCR\u9175\u7d20\u63a8\u5968\u306e\u53cd\u5fdc\u7d44\u6210\u30fb\u53cd\u5fdc\u6761\u4ef6\u3067\u8abf\u88fd\u3057\u3066\u304f\u3060\u3055\u3044\u3002<\/p>\n<div id=\"sosei\">\n<p>\u25cf\u53cd\u5fdc\u7d44\u6210<\/p>\n<dl>\n<dt>10x Ex buffer<\/dt>\n<dd>2.0 ul<\/dd>\n<dt>dNTPs (each 2.5 mM)<\/dt>\n<dd>1.6 ul<\/dd>\n<dt>10 uM Forward primer<\/dt>\n<dd>1.0 ul<\/dd>\n<dt>10 uM Reverse primer<\/dt>\n<dd>1.0 ul<\/dd>\n<dt>Template DNA (2 ng)<\/dt>\n<dd>4.0ul<\/dd>\n<dt>ExTaq HS[TaKaRa]\uff085 U\/ul)<\/dt>\n<dd>0.15 ul<\/dd>\n<dt class=\"underline\">\u6ec5\u83cc\u84b8\u7559\u6c34<\/dt>\n<dd class=\"underline\">10.25 ul<\/dd>\n<dt>Total<\/dt>\n<dd>20.0 ul<\/dd>\n<\/dl>\n<\/div>\n<div id=\"ondo\">\n<p>&nbsp;<\/p>\n<p>\u25cf\u53cd\u5fdc\u6e29\u5ea6<\/p>\n<dl>\n<dt>94\u00b0C<\/dt>\n<dd>2min<\/dd>\n<dt>94\u00b0C<\/dt>\n<dd>20sec\uff5c<\/dd>\n<dt>65\u00b0C<\/dt>\n<dd>15sec\uff5c35 cycles<\/dd>\n<dt>72\u00b0C<\/dt>\n<dd>20sec\uff5c<\/dd>\n<dt>72\u00b0C<\/dt>\n<dd>5min<\/dd>\n<\/dl>\n<\/div>\n","protected":false},"excerpt":{"rendered":"\u25cf\u30d7\u30e9\u30a4\u30de\u30fc\u914d\u5217\uff085\u2019\u21923\u2019\uff09\/\u30a2\u30f3\u30d7\u30ea\u30b3\u30f3\u30b7\u30fc\u30b1\u30f3\u30b9\u89e3\u6790\uff08illumina MiSeq\uff09\u7528 1st-cMiFish F\uff1a \u2460ACACTCTTTCCCTACACGACGCTCTTCCGATCT-NNNNNN&#8211; [&hellip;]","protected":false},"author":1,"featured_media":0,"parent":972,"menu_order":10,"comment_status":"closed","ping_status":"closed","template":"","meta":{"footnotes":""},"class_list":["post-5374","page","type-page","status-publish","hentry"],"_links":{"self":[{"href":"https:\/\/gikenbio.com\/2025test\/wp-json\/wp\/v2\/pages\/5374","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/gikenbio.com\/2025test\/wp-json\/wp\/v2\/pages"}],"about":[{"href":"https:\/\/gikenbio.com\/2025test\/wp-json\/wp\/v2\/types\/page"}],"author":[{"embeddable":true,"href":"https:\/\/gikenbio.com\/2025test\/wp-json\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"https:\/\/gikenbio.com\/2025test\/wp-json\/wp\/v2\/comments?post=5374"}],"version-history":[{"count":9,"href":"https:\/\/gikenbio.com\/2025test\/wp-json\/wp\/v2\/pages\/5374\/revisions"}],"predecessor-version":[{"id":6638,"href":"https:\/\/gikenbio.com\/2025test\/wp-json\/wp\/v2\/pages\/5374\/revisions\/6638"}],"up":[{"embeddable":true,"href":"https:\/\/gikenbio.com\/2025test\/wp-json\/wp\/v2\/pages\/972"}],"wp:attachment":[{"href":"https:\/\/gikenbio.com\/2025test\/wp-json\/wp\/v2\/media?parent=5374"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}