{"id":1987,"date":"2014-09-17T10:33:26","date_gmt":"2014-09-17T01:33:26","guid":{"rendered":"http:\/\/jcm.brc.riken.jp\/ja\/?page_id=1987"},"modified":"2017-03-24T14:43:45","modified_gmt":"2017-03-24T05:43:45","slug":"quality_control","status":"publish","type":"page","link":"http:\/\/jcm.brc.riken.jp\/ja\/quality_control","title":{"rendered":"\u5fae\u751f\u7269\u682a\u306e\u54c1\u8cea\u7ba1\u7406"},"content":{"rendered":"<p>\u5fae\u751f\u7269\u6750\u6599\u958b\u767a\u5ba4 (JCM) \u3067\u306f\u3001\u4ee5\u4e0b\u306b\u793a\u3059\u691c\u67fb\u3092\u5fae\u751f\u7269\u682a\u53d7\u5165\u6642\u4e26\u3073\u306b\u4fdd\u5b58\u6a19\u54c1\u4f5c\u88fd\u6642\u306b\u5171\u901a\u3057\u3066\u5b9f\u65bd\u3057\u3066\u3044\u307e\u3059\u3002\u307e\u305f\u3001\u5fc5\u8981\u306b\u5fdc\u3058\u3066\u65e2\u4fdd\u5b58\u682a\u306e\u54c1\u8cea\u691c\u67fb\u3082\u5b9f\u65bd\u3057\u3066\u3044\u307e\u3059\u3002\u306a\u304a\u3001\u30aa\u30f3\u30e9\u30a4\u30f3\u30ab\u30bf\u30ed\u30b0\u306b\u63b2\u8f09\u3057\u3066\u3044\u308b\u7279\u6027\u3084\u6a5f\u80fd\u306f\u6587\u732e\u60c5\u5831\u7b49\u306b\u57fa\u3065\u304f\u3082\u306e\u3067\u3001\u305d\u308c\u3089\u3092\u4fdd\u8a3c\u3059\u308b\u3082\u306e\u3067\u306f\u3042\u308a\u307e\u305b\u3093\u3002\u7814\u7a76\u306b\u5fc5\u8981\u306a\u7279\u6027\u7b49\u306b\u3064\u3044\u3066\u306f\u3001\u5229\u7528\u8005\u3054\u81ea\u8eab\u3067\u3054\u78ba\u8a8d\u304f\u3060\u3055\u3044\u3002<\/p>\n<ul type=\"disc\">\n<li style=\"height: 25px;\">\u5b9f\u65bd\u9805\u76ee<\/li>\n<ul type=\"circle\" style=\"margin:0 0 0 20px; padding:0;\">\n<li style=\"height: 10px;\">\u53d7\u5165\u6642\u691c\u67fb<\/li>\n<blockquote style=\"color:#000000; background-color: #ffffff; border:0px;margin:0 50px 0 20px; padding: 0px;\"><p>\n\u65b0\u898f\u306b\u5bc4\u8a17\u307e\u305f\u306f\u8b72\u6e21\u3055\u308c\u305f\u5fae\u751f\u7269\u682a\u3092\u5fae\u751f\u7269\u682a\u62c5\u5f53\u8005\u304c\u57f9\u990a\u3057\u3001\u751f\u6b8b\u6027\u3001\u30b3\u30ed\u30cb\u30fc\u5f62\u614b\u3001\u7d30\u80de\u5f62\u614b\u3001\u30b3\u30f3\u30bf\u30df\u30cd\u30fc\u30b7\u30e7\u30f3\u306e\u6709\u7121\u306a\u3069\u3092\u78ba\u8a8d\u3057\u307e\u3059\u3002 \u540c\u6642\u306b PCR \u306b\u3066 rRNA \u907a\u4f1d\u5b50\u7b49\u306e\u907a\u4f1d\u5b50\u9818\u57df\u3092\u5897\u5e45\u3057\u3066\u5869\u57fa\u914d\u5217\u3092\u6c7a\u5b9a\u3057\u3001\u5bc4\u8a17\u8005\u3088\u308a\u63d0\u793a\u3055\u308c\u305f\u5869\u57fa\u914d\u5217\u3084\u516c\u958b\u5869\u57fa\u914d\u5217\u3068\u306e\u540c\u4e00\u6027\u3092\u78ba\u8a8d\u3059\u308b<a href=\"#dna\">\u907a\u4f1d\u5b50\u691c\u67fb<\/a>\u3092\u884c\u3044\u307e\u3059\u3002\u306a\u304a\u3001\u907a\u4f1d\u5b50\u691c\u67fb\u306f2011\u5e74\u5ea6\u4ee5\u964d\u306b\u5bc4\u8a17\u3055\u308c\u305f\u5fae\u751f\u7269\u682a\u304c\u5bfe\u8c61\u3067\u3059\u30022015\u5e74\u4ee5\u964d\u306b\u5bc4\u8a17\u3055\u308c\u305f\u30a2\u30fc\u30ad\u30a2\u682a\u306b\u3064\u3044\u3066\u306f\u3001\u7d30\u83cc\u306e16S rRNA\u907a\u4f1d\u5b50\u3092\u5897\u5e45\u3059\u308bPCR\u89e3\u6790\u306b\u3088\u308a\u3001\u7d30\u83cc\u306e\u6df7\u5165\u306e\u6709\u7121\u3082\u691c\u67fb\u3057\u3066\u3044\u307e\u3059\u3002\n<\/blockquote>\n<li style=\"height: 10px;\">\u4fdd\u5b58\u6a19\u54c1\u4f5c\u88fd\u6642\u691c\u67fb<\/li>\n<blockquote style=\"color:#000000; background-color: #ffffff; border:0px;margin:0 50px 0 20px; padding: 0px;\"><p>\n\u51cd\u7d50\u4fdd\u5b58\u3001\u51cd\u7d50\u4e7e\u71e5\u4fdd\u5b58\u3001L-\u4e7e\u71e5\u4fdd\u5b58\u306a\u3069\u306e\u65b9\u6cd5\u3067\u4f5c\u88fd\u3055\u308c\u305f\u6a19\u54c1\u306b\u3064\u3044\u3066\u3001\u5fae\u751f\u7269\u682a\u306e\u751f\u6b8b\u6027\u3068\u30b3\u30f3\u30bf\u30df\u30cd\u30fc\u30b7\u30e7\u30f3\u306e\u6709\u7121\u3092\u78ba\u8a8d\u3057\u307e\u3059\u3002\n<\/p><\/blockquote>\n<\/ul>\n<\/ul>\n<ul type=\"disc\">\n<li style=\"height: 10px;\">\u672a\u5b9f\u65bd\u9805\u76ee<\/li>\n<blockquote style=\"color:#000000; background-color: #ffffff; border:0px;margin:0 50px 0 20px; padding: 0px;\"><p>\n\u30aa\u30f3\u30e9\u30a4\u30f3\u30ab\u30bf\u30ed\u30b0\u306b\u63b2\u8f09\u3057\u3066\u3044\u308b\u7279\u6027\u3084\u6a5f\u80fd\u306b\u3064\u3044\u3066\u306f\u3001\u691c\u67fb\u3092\u5b9f\u65bd\u3057\u3066\u3044\u307e\u305b\u3093\u3002\n<\/p><\/blockquote>\n<\/ul>\n<ul type=\"disc\">\n<li style=\"height: 10px;\">\u8ffd\u52a0\u5b9f\u65bd\u9805\u76ee<\/li>\n<blockquote style=\"color:#000000; background-color: #ffffff; border:0px;margin:0 50px 0 20px; padding: 0px;\"><p>\n\u4e0a\u8a18\u306e\u5b9f\u65bd\u9805\u76ee\u4ee5\u5916\u306e\u691c\u67fb\u3092\u5b9f\u65bd\u3057\u305f\u5fae\u751f\u7269\u682a\u306b\u3064\u3044\u3066\u306f\u3001\u691c\u67fb\u7d50\u679c\u3092\u30ab\u30bf\u30ed\u30b0\u306b\u8a18\u8f09\u3057\u3066\u3044\u307e\u3059\u3002\n<\/p><\/blockquote>\n<\/ul>\n<p><a name=\"dna\"><\/a><br \/>\n\u907a\u4f1d\u5b50\u691c\u67fb\u306e\u65b9\u6cd5<\/p>\n<blockquote style=\"color:#000000; background-color: #ffffff; border:0px;margin:0 50px -10px 20px; padding: 0px;\"><p>\n\u5fae\u751f\u7269\u6750\u6599\u958b\u767a\u5ba4\u3067\u5b9f\u65bd\u3057\u3066\u3044\u308b\u6a19\u6e96\u7684\u306a\u907a\u4f1d\u5b50\u691c\u67fb\u306e\u624b\u9806\u306f\u3001\u4ee5\u4e0b\u306e\u901a\u308a\u3067\u3059\u3002\n<\/p><\/blockquote>\n<blockquote style=\"color:#000000; background-color: #ffffff; border:0px;margin:0 60px 0 40px; padding: 0px;\"><p>\n(1) \u83cc\u4f53\u7834\u7815\u6db2\u306e\u8abf\u88fd<br \/>\n(2) PCR<br \/>\n(3) \u30b7\u30fc\u30af\u30a8\u30f3\u30b9\u89e3\u6790\n<\/blockquote>\n<blockquote style=\"color:#000000; background-color: #ffffff; border:0px;margin:0 50px -10px 20px; padding: 0px;\"><p>\n\u4f7f\u7528\u3059\u308b\u3082\u306e\n<\/p><\/blockquote>\n<blockquote style=\"color:#000000; background-color: #ffffff; border:0px;margin:0 60px -10px 40px; padding: 0px;\"><p>\n&#8226; \u83cc\u4f53\u7834\u7815\u88c5\u7f6e\u3068\u7834\u7815\u7528\u6d88\u8017\u54c1\n<\/p><\/blockquote>\n<blockquote style=\"color:#000000; background-color: #ffffff; border:0px;margin:0 70px 0 60px; padding: 0px;\"><p>\nFastPrep-24 (MP Biomedicals)<br \/>\n\u30ac\u30e9\u30b9\u30d3\u30fc\u30ba (SIGMA acid-washed glass beads G4649 [\u2266106 &#181;m] \u7b49) \u4e7e\u71b1\u6ec5\u83cc<br \/>\n2.0 mL \u30b9\u30af\u30ea\u30e5\u30fc\u30ad\u30e3\u30c3\u30d7\u30c1\u30e5\u30fc\u30d6\uff08\u30a2\u30ba\u30ef\u30f3 C-2.0 \u306a\u3069\uff09\n<\/blockquote>\n<blockquote style=\"color:#000000; background-color: #ffffff; border:0px;margin:0 60px -10px 40px; padding: 0px;\"><p>\n&#8226; DNA \u7cbe\u88fd\u30ad\u30c3\u30c8\n<\/p><\/blockquote>\n<blockquote style=\"color:#000000; background-color: #ffffff; border:0px;margin:0 70px 0 60px; padding: 0px;\"><p>\nMultiScreen PCR96 \u30d5\u30a3\u30eb\u30bf\u30fc\u30d7\u30ec\u30fc\u30c8 (\u30e1\u30eb\u30af\u30df\u30ea\u30dd\u30a2 MSNU03010)<br \/>\nMonoFas DNA \u7cbe\u88fd\u30ad\u30c3\u30c8I (GL Sciences)\n<\/blockquote>\n<blockquote style=\"color:#000000; background-color: #ffffff; border:0px;margin:0 60px -10px 40px; padding: 0px;\"><p>\n&#8226; PCR\n<\/p><\/blockquote>\n<blockquote style=\"color:#000000; background-color: #ffffff; border:0px;margin:0 70px 0 60px; padding: 0px;\"><p>\n<i>TaKaRa Ex Taq<\/i><sup>&#174;<\/sup> Hot Start Version (\u30bf\u30ab\u30e9\u30d0\u30a4\u30aa)<br \/>\n\u305d\u306e\u4ed6\u3001\u907a\u4f1d\u5b50\u89e3\u6790\u306b\u5fc5\u8981\u306a\u4e00\u822c\u7684\u306a\u8a66\u85ac\u3001\u7de9\u885d\u6db2\u3001\u5668\u5177\n<\/blockquote>\n<blockquote style=\"color:#000000; background-color: #ffffff; border:0px;margin:0 50px -10px 20px; padding: 0px;\"><p>\n(1) \u83cc\u4f53\u7834\u7815\u6db2\u306e\u8abf\u88fd\n<\/p><\/blockquote>\n<ol style=\"margin:0 50px 0 20px;\">\n<li>2.0 mL \u30b9\u30af\u30ea\u30e5\u30fc\u30ad\u30e3\u30c3\u30d7\u30de\u30a4\u30af\u30ed\u30c1\u30e5\u30fc\u30d6\uff08\u83cc\u4f53\u7834\u7815\u88c5\u7f6e\u3067\u4f7f\u7528\u3067\u304d\u308b\u3082\u306e\uff09\u306b\u3001\u304a\u3088\u305d 0.3 g \u306e\u30ac\u30e9\u30b9\u30d3\u30fc\u30ba\u3068 400 &#181;L \u306e TE\u30d0\u30c3\u30d5\u30a1\u30fc\u3092\u5165\u308c\u3001\u9069\u91cf\u306e\u83cc\u4f53\u3092\u61f8\u6fc1\u3059\u308b\u3002<\/li>\n<li>FastPrep-24 \u3092\u7528\u3044\u3066\u83cc\u4f53\u3092\u7834\u7815\u3059\u308b\u3002\u901f\u5ea66\u3001\u6642\u959320\u79d2\u3092\u57fa\u672c\u3068\u3059\u308b\u3002<\/li>\n<li>\u9060\u5fc3\u5206\u96e2\u306b\u3088\u308a\u83cc\u4f53\u7834\u7815\u6b8b\u6e23\u3092\u6c88\u6bbf\u3055\u305b\u305f\u4e0a\u6e05\u3092\u65b0\u3057\u3044\u30de\u30a4\u30af\u30ed\u30c1\u30e5\u30fc\u30d6\u306b\u79fb\u3057\u3001\u83cc\u4f53\u7834\u7815\u6db2\u3068\u3059\u308b\u3002<\/li>\n<li>\u5fc5\u8981\u306b\u5fdc\u3058\u3066\u83cc\u4f53\u7834\u7815\u6db2\u3092 MonoFas \u3067\u7cbe\u88fd\u3001\u6fc3\u7e2e\u3059\u308b\u3002<\/li>\n<\/ol>\n<blockquote style=\"color:#000000; background-color: #ffffff; border:0px;margin:0 50px -10px 20px; padding: 0px;\"><p>\n(2) PCR\n<\/p><\/blockquote>\n<ol style=\"margin:0 50px 0 20px;\">\n<li>\u83cc\u4f53\u7834\u7815\u6db2\u307e\u305f\u306f\u305d\u306e\u7cbe\u88fd\u6eb6\u6db2\u3092\u9069\u5b9c\u5e0c\u91c8\u3057\u305f\u3082\u306e\u3092\u92f3\u578b\u3068\u3059\u308b\u3002<\/li>\n<li>\u88681\u301c4 \u306b\u793a\u3059 PCR \u53cd\u5fdc\u6eb6\u6db2\u53ca\u3073\u30b5\u30a4\u30af\u30eb\u6761\u4ef6\u306b\u3066\u3001\u6a19\u7684\u907a\u4f1d\u5b50\u9818\u57df\u3092\u5897\u5e45\u3059\u308b\u3002<\/li>\n<li>PCR \u7523\u7269\u3092\u96fb\u6c17\u6cf3\u52d5\u3057\u3001\u30b7\u30fc\u30b1\u30f3\u30b9\u89e3\u6790\u306b\u9069\u3057\u305f\u5897\u5e45\u7523\u7269\u304c\u5f97\u3089\u308c\u3066\u3044\u308b\u3053\u3068\u3092\u78ba\u8a8d\u3059\u308b\u3002<\/li>\n<\/ol>\n<blockquote style=\"color:#000000; background-color: #ffffff; border:0px;margin:0 50px -10px 20px; padding: 0px;\"><p>\n(3) \u30b7\u30fc\u30af\u30a8\u30f3\u30b9\u89e3\u6790\n<\/p><\/blockquote>\n<ol style=\"margin:0 60px 0 20px;\">\n<li>MultiScreen \u3084 MonoFas \u7b49\u306e DNA \u7cbe\u88fd\u30ad\u30c3\u30c8\u306b\u3088\u308a\u3001PCR \u7523\u7269\u3092\u7cbe\u88fd\u3059\u308b\u3002<\/li>\n<li>\u30b7\u30fc\u30b1\u30f3\u30b5\u30fc\u6307\u5b9a\u306e\u624b\u9806\u306b\u5f93\u3044\u3001PCR \u7523\u7269\u306e\u5869\u57fa\u914d\u5217\u3092\u6c7a\u5b9a\u3059\u308b\u3002\u30b7\u30fc\u30b1\u30f3\u30b9\u30d7\u30e9\u30a4\u30de\u30fc\u306f\u88682\u306b\u793a\u3059\u3082\u306e\u304b\u3089\u9069\u5b9c\u9078\u629e\u3059\u308b\u3002<\/li>\n<li>\u30b7\u30fc\u30b1\u30f3\u30b9\u30c7\u30fc\u30bf\u306f\u5fc5\u8981\u306a\u4fee\u6b63\u3092\u52a0\u3048\u3001\u5bc4\u8a17\u8005\u3088\u308a\u63d0\u793a\u3055\u308c\u305f\u914d\u5217\u30c7\u30fc\u30bf\u3068\u6bd4\u8f03\u3059\u308b\u3002\u5fc5\u8981\u306b\u5fdc\u3058\u3066\u3001\u5869\u57fa\u914d\u5217\u30c7\u30fc\u30bf\u30d9\u30fc\u30b9\u4e0a\u306e\u914d\u5217\u30c7\u30fc\u30bf\u3068\u6bd4\u8f03\u3059\u308b\u3002<\/li>\n<\/ol>\n<p>&nbsp;<br \/>\nTable 1. PCR mixture<\/p>\n<table width=\"300\" border=\"0\" cellspacing=\"0\" cellpadding=\"0\">\n<tbody>\n<tr bgcolor=\"gray\">\n<th height=\"25\">Reagent<\/th>\n<th style=\"text-align: center\">Volume (&#181;L)<\/th>\n<\/tr>\n<tr>\n<td height=\"25\">Milli-Q Water<\/td>\n<td style=\"text-align: right\">37.75<\/td>\n<\/tr>\n<tr>\n<td height=\"25\">10 x Buffer<\/td>\n<td style=\"text-align: right\">5.0<\/td>\n<\/tr>\n<tr>\n<td height=\"25\">dNTPs (2.5 mM)<\/td>\n<td style=\"text-align: right\">4.0<\/td>\n<\/tr>\n<tr>\n<td height=\"25\">Forward primer (10 &#181;M)<\/td>\n<td style=\"text-align: right\">1.0<\/td>\n<\/tr>\n<tr>\n<td height=\"25\">Reverse primer (10 &#181;M)<\/td>\n<td style=\"text-align: right\">1.0<\/td>\n<\/tr>\n<tr>\n<td height=\"25\"><i>TaKaRa Ex Taq<\/i><sup>&#174;<\/sup> HS<\/td>\n<td style=\"text-align: right\">0.25<\/td>\n<\/tr>\n<tr>\n<td height=\"25\">Template DNA<\/td>\n<td style=\"text-align: right\">1.0<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<p>Total reaction volume 50 &#181;L.<\/p>\n<p>&nbsp;<br \/>\nTable 2. Primers used in PCR amplificaion and sequencing<\/p>\n<table width=\"600\" border=\"0\" cellspacing=\"0\" cellpadding=\"0\">\n<tbody>\n<tr>\n<th style=\"text-align: center\" height=\"25\">Primer<\/th>\n<th style=\"text-align: center\">Sequence (5&#8217;\u20133&#8242;)<\/th>\n<th style=\"text-align: center\">Target Group<\/th>\n<\/tr>\n<tr>\n<td style=\"text-align: center\" height=\"25\">27F<sup>1<\/sup><\/td>\n<td style=\"text-align: center\">AGAGTTTGATCCTGGCTCAG<\/td>\n<td style=\"text-align: center\">Bacteria<\/td>\n<\/tr>\n<tr>\n<td style=\"text-align: center\" height=\"25\">786F<sup>2<\/sup><\/td>\n<td style=\"text-align: center\">GATTAGATACCCTGGTAG<\/td>\n<td style=\"text-align: center\">Bacteria<\/td>\n<\/tr>\n<tr>\n<td style=\"text-align: center\" height=\"25\">1492R<sup>3<\/sup><\/td>\n<td style=\"text-align: center\">GGTTACCTTGTTACGACTT<\/td>\n<td style=\"text-align: center\">Bacteria<\/td>\n<\/tr>\n<tr>\n<td style=\"text-align: center\" height=\"25\">907R<sup>1<\/sup><\/td>\n<td style=\"text-align: center\">CCGTCAATTCMTTTRAGTTT<\/td>\n<td style=\"text-align: center\">Bacteria<\/td>\n<\/tr>\n<tr>\n<td style=\"text-align: center\" height=\"25\">826R<sup>4<\/sup><\/td>\n<td style=\"text-align: center\">GACTACCAGGGTATCTAATCC<\/td>\n<td style=\"text-align: center\">Bacteria<\/td>\n<\/tr>\n<tr>\n<td style=\"text-align: center\" height=\"25\">519R<sup>1<\/sup><\/td>\n<td style=\"text-align: center\">GWATTACCGCGGCKGCTG<\/td>\n<td style=\"text-align: center\">Bacteria\/Archaea<\/td>\n<\/tr>\n<tr>\n<td style=\"text-align: center\" height=\"25\">530F<sup>1<\/sup><\/td>\n<td style=\"text-align: center\">GTGCCAGCMGCCGCGG<\/td>\n<td style=\"text-align: center\">Bacteria\/Archaea<\/td>\n<\/tr>\n<tr>\n<td style=\"text-align: center\" height=\"25\">1053F<sup>2<\/sup><\/td>\n<td style=\"text-align: center\">GCATGGCYGYCGTCAG<\/td>\n<td style=\"text-align: center\">Bacteria\/Archaea<\/td>\n<\/tr>\n<tr>\n<td style=\"text-align: center\" height=\"25\">A21F<sup>5<\/sup><\/td>\n<td style=\"text-align: center\">TTCCGGTTGATCCYGCCGGA<\/td>\n<td style=\"text-align: center\">Archaea<\/td>\n<\/tr>\n<tr>\n<td style=\"text-align: center\" height=\"25\">A348F<sup>6<\/sup><\/td>\n<td style=\"text-align: center\">TCCAGGCCCTACGGG<\/td>\n<td style=\"text-align: center\">Archaea<\/td>\n<\/tr>\n<tr>\n<td style=\"text-align: center\" height=\"25\">A1400R<sup>7<\/sup><\/td>\n<td style=\"text-align: center\">GACGGGCGGTGTGTGC<\/td>\n<td style=\"text-align: center\">Archaea<\/td>\n<\/tr>\n<tr>\n<td style=\"text-align: center\" height=\"25\">A1100R<sup>7<\/sup><\/td>\n<td style=\"text-align: center\">CGGGTCTCGCTCGTT<\/td>\n<td style=\"text-align: center\">Archaea<\/td>\n<\/tr>\n<tr>\n<td style=\"text-align: center\" height=\"25\">ITS5<sup>8<\/sup><\/td>\n<td style=\"text-align: center\">GGAAGTAAAAGTCGTAACAAGG<\/td>\n<td style=\"text-align: center\">Fungi<\/td>\n<\/tr>\n<tr>\n<td style=\"text-align: center\" height=\"25\">ITS3<sup>8<\/sup><\/td>\n<td style=\"text-align: center\">GCATCGATGAAGAACGCAGC<\/td>\n<td style=\"text-align: center\">Fungi<\/td>\n<\/tr>\n<tr>\n<td style=\"text-align: center\" height=\"25\">NL1<sup>9<\/sup><\/td>\n<td style=\"text-align: center\">GCATATCAATAAGCGGAGGAAAAG<\/td>\n<td style=\"text-align: center\">Fungi<\/td>\n<\/tr>\n<tr>\n<td style=\"text-align: center\" height=\"25\">NL4<sup>9<\/sup><\/td>\n<td style=\"text-align: center\">GGTCCGTGTTTCAAGACGG<\/td>\n<td style=\"text-align: center\">Fungi<\/td>\n<\/tr>\n<tr>\n<td style=\"text-align: center\" height=\"25\">ITS4<sup>8<\/sup><\/td>\n<td style=\"text-align: center\">TCCTCCGCTTATTGATATGC<\/td>\n<td style=\"text-align: center\">Fungi<\/td>\n<\/tr>\n<tr>\n<td style=\"text-align: center\" height=\"25\">ITS2<sup>8<\/sup><\/td>\n<td style=\"text-align: center\">GCTGCGTTCTTCATCGATGC<\/td>\n<td style=\"text-align: center\">Fungi<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<p>&nbsp;<br \/>\nTable 3. PCR primer sets<\/p>\n<table width=\"350\" border=\"0\" cellspacing=\"0\" cellpadding=\"0\">\n<tbody>\n<tr>\n<th style=\"text-align: center\" height=\"25\">Target Group<\/th>\n<th>Primer set<\/th>\n<th>Target gene<\/th>\n<\/tr>\n<tr>\n<td style=\"text-align: center\" height=\"25\">Bacteria<\/td>\n<td>27F + 1492R<\/td>\n<td>16S rRNA<\/td>\n<\/tr>\n<tr>\n<td style=\"text-align: center\" height=\"25\">Archaea<\/td>\n<td>A21F + A1100R<\/td>\n<td>16S rRNA<\/td>\n<\/tr>\n<tr>\n<td style=\"text-align: center\" height=\"25\">Archaea<\/td>\n<td>A21F + A1400R<\/td>\n<td>16S rRNA<\/td>\n<\/tr>\n<tr>\n<td style=\"text-align: center\" height=\"25\">Fungi<\/td>\n<td>ITS5 + NL4<\/td>\n<td>ITS, LSU<\/td>\n<\/tr>\n<tr>\n<td style=\"text-align: center\" height=\"25\">Fungi<\/td>\n<td>ITS5 + ITS4<\/td>\n<td>ITS<\/td>\n<\/tr>\n<tr>\n<td style=\"text-align: center\" height=\"25\">Fungi<\/td>\n<td>NL1 + NL4<\/td>\n<td>LSU<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<p>&nbsp;<br \/>\nTable 4. PCR cycling condition<\/p>\n<table width=\"300\" border=\"0\" cellspacing=\"0\" cellpadding=\"0\">\n<tbody>\n<tr>\n<th style=\"text-align: center\" height=\"25\">Step<\/th>\n<th style=\"text-align: center\">Temp. (&#176;C)<\/th>\n<th style=\"text-align: center\">Time<\/th>\n<\/tr>\n<tr>\n<td style=\"text-align: center\" height=\"25\">1<\/td>\n<td style=\"text-align: center\">98<\/td>\n<td style=\"text-align: center\">2 min.<\/td>\n<\/tr>\n<tr>\n<td style=\"text-align: center\" height=\"25\">2<\/td>\n<td style=\"text-align: center\">98<\/td>\n<td style=\"text-align: center\">10 sec.<\/td>\n<\/tr>\n<tr>\n<td style=\"text-align: center\" height=\"25\">3<\/td>\n<td style=\"text-align: center\">53<\/td>\n<td style=\"text-align: center\">20 sec.<\/td>\n<\/tr>\n<tr>\n<td style=\"text-align: center\" height=\"25\">4<\/td>\n<td style=\"text-align: center\">72<\/td>\n<td style=\"text-align: center\">60 sec.<\/td>\n<\/tr>\n<tr>\n<td style=\"text-align: center\" height=\"25\">5<\/td>\n<td style=\"text-align: center\" colspan=\"2\">Followed steps 2-4 by 30 to 35 cycles<\/td>\n<\/tr>\n<tr>\n<td style=\"text-align: center\" height=\"25\">6<\/td>\n<td style=\"text-align: center\">72<\/td>\n<td style=\"text-align: center\">5 min.<\/td>\n<\/tr>\n<tr>\n<td style=\"text-align: center\" height=\"25\">7<\/td>\n<td style=\"text-align: center\">10<\/td>\n<td style=\"text-align: center\">hold<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<p>&nbsp;<br \/>\n<strong>Reference<\/strong><\/p>\n<ol>\n<li>Lane, D. J. (1991). 16S\/23S rRNA sequencing, <em>In:<\/em> Stackebrandt, E. and Goodfellow, M. (eds.), Nucleic Acid Techniques in Bacterial Systematics, pp. 115\u2013176, Jon Wiley &#038; Sons, Chichester.<\/li>\n<li>Baker, G. C., Smith, J. J., and Cowan, D. A. (2003). Review and re-analysis of domain-specific 16S primers. J. Microbiol. Methods <strong>55:<\/strong> 541\u2013555.<\/li>\n<li>Turner, S., Pryer, K. M., Miao, V. P., and Palmer, J. D. (1999). Investigating deep phylogenetic relationships among cyanobacteria and plastids by small subunit rRNA sequence analysis. J. Euk. Microbiol. <strong>46:<\/strong> 327\u2013338.<\/li>\n<li>Youssef, N., Sheik, C. S., Krumholz, L. R., Najar, F. Z., Roe, B. A., and Elshahed, M. S. (2009). Comparison of species richness estimates obtained using nearly complete fragments and simulated pyrosequencing-generated fragments in 16S rRNA gene-based environmental surveys. Appl. Environ. Microbiol. <strong>75:<\/strong> 5227\u20135236.<\/li>\n<li>DeLong, E. F. (1992). Archaea in coastal marine environments. Proc. Natl. Acad. Sci. USA <strong>89:<\/strong> 5685\u20135689.<\/li>\n<li>Achenbach, L. and Woese, C. (1995). Appendix 6. 16S and 23S rRNA-like primers, <em>In:<\/em> Robb, F. T. and Place, A. R. (eds.), Archaea: a Laboratory Manual. Thermophiles, pp. 201-203, Cold Spring Harbor Laboratory Press, New York.<\/li>\n<li>Itoh, T., Suzuki, K., and Nakase, T. (1998). Occurrence of introns in the 16S rRNA genes of members of the genus Thermoproteus. Arch. Microbiol. <strong>170:<\/strong> 155\u2013161.<\/li>\n<li>White, T. J., Bruns, T., Lee, S., and Taylor, J. W. (1990). Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics, In: Innis, M. A., Gelfand, D. H., Sninsky, J. J., and White, T. J. (eds.), PCR Protocols: A Guide to Methods and Applications, pp. 315-322, Academic Press, New York.<\/li>\n<li>O\u2019Donell, K. (1993). <i>Fusarium<\/i> and its near relatives, <em>In:<\/em> Reynolds, D. R. and Taylor, J. W. (eds.), The Fungal Holomorph: Mitotic, Meiotic and Pleomorphic Speciation in Fungal Systematics, pp. 225-233, CAB International, Wallingford.<\/li>\n<\/ol>\n","protected":false},"excerpt":{"rendered":"<p>\u5fae\u751f\u7269\u6750\u6599\u958b\u767a\u5ba4 (JCM) \u3067\u306f\u3001\u4ee5\u4e0b\u306b\u793a\u3059\u691c\u67fb\u3092\u5fae\u751f\u7269\u682a\u53d7\u5165\u6642\u4e26\u3073\u306b\u4fdd\u5b58\u6a19\u54c1\u4f5c\u88fd\u6642\u306b\u5171\u901a\u3057\u3066\u5b9f\u65bd\u3057\u3066\u3044\u307e\u3059\u3002\u307e\u305f\u3001\u5fc5\u8981\u306b\u5fdc\u3058\u3066\u65e2\u4fdd\u5b58\u682a\u306e\u54c1\u8cea\u691c\u67fb\u3082\u5b9f\u65bd\u3057\u3066\u3044\u307e\u3059\u3002\u306a\u304a\u3001\u30aa\u30f3\u30e9\u30a4\u30f3\u30ab\u30bf\u30ed\u30b0\u306b\u63b2\u8f09\u3057\u3066\u3044\u308b\u7279\u6027\u3084\u6a5f\u80fd\u306f\u6587\u732e\u60c5\u5831\u7b49\u306b\u57fa\u3065\u304f\u3082\u306e\u3067\u3001\u305d\u308c\u3089\u3092\u4fdd\u8a3c\u3059\u308b\u3082\u306e\u3067\u306f\u3042\u308a\u307e\u305b\u3093\u3002\u7814\u7a76\u306b\u5fc5\u8981\u306a\u7279\u6027\u7b49\u306b\u3064\u3044\u3066\u306f\u3001\u5229\u7528\u8005\u3054\u81ea\u8eab\u3067\u3054\u78ba\u8a8d\u304f\u3060\u3055\u3044\u3002 \u5b9f\u65bd\u9805\u76ee \u53d7\u5165\u6642\u691c\u67fb \u65b0\u898f\u306b\u5bc4\u8a17\u307e\u305f\u306f\u8b72\u6e21\u3055\u308c\u305f\u5fae\u751f\u7269\u682a\u3092\u5fae [&hellip;]<\/p>\n","protected":false},"author":16,"featured_media":0,"parent":0,"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-1987","page","type-page","status-publish","hentry"],"_links":{"self":[{"href":"http:\/\/jcm.brc.riken.jp\/ja\/wp-json\/wp\/v2\/pages\/1987","targetHints":{"allow":["GET"]}}],"collection":[{"href":"http:\/\/jcm.brc.riken.jp\/ja\/wp-json\/wp\/v2\/pages"}],"about":[{"href":"http:\/\/jcm.brc.riken.jp\/ja\/wp-json\/wp\/v2\/types\/page"}],"author":[{"embeddable":true,"href":"http:\/\/jcm.brc.riken.jp\/ja\/wp-json\/wp\/v2\/users\/16"}],"replies":[{"embeddable":true,"href":"http:\/\/jcm.brc.riken.jp\/ja\/wp-json\/wp\/v2\/comments?post=1987"}],"version-history":[{"count":9,"href":"http:\/\/jcm.brc.riken.jp\/ja\/wp-json\/wp\/v2\/pages\/1987\/revisions"}],"predecessor-version":[{"id":2953,"href":"http:\/\/jcm.brc.riken.jp\/ja\/wp-json\/wp\/v2\/pages\/1987\/revisions\/2953"}],"wp:attachment":[{"href":"http:\/\/jcm.brc.riken.jp\/ja\/wp-json\/wp\/v2\/media?parent=1987"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}