{"id":3434,"date":"2013-04-10T16:44:29","date_gmt":"2013-04-10T07:44:29","guid":{"rendered":"http:\/\/cell.brc.riken.jp:7321\/?page_id=3434"},"modified":"2016-12-15T10:02:02","modified_gmt":"2016-12-15T01:02:02","slug":"str","status":"publish","type":"page","link":"http:\/\/cell.brc.riken.jp\/ja\/quality\/str","title":{"rendered":"STR"},"content":{"rendered":"<p>\u30d2\u30c8\u7d30\u80de\u6750\u6599\u306e\u7531\u6765\u500b\u4f53\u306e\u8b58\u5225\u540c\u5b9a\u3001\u8d77\u6e90\u306e\u78ba\u8a8d\u3001\u30af\u30ed\u30b9\u30b3\u30f3\u30bf\u30df\u30cd\u30fc\u30b7\u30e7\u30f3\u306a\u3069\u3092\u78ba\u8a8d\u3059\u308b\u305f\u3081\u306b\u3001Short Tandem Repeat (STR) \u591a\u578b\u89e3\u6790\u3092\u884c\u3063\u3066\u3044\u307e\u3059\u3002\u3053\u306e\u691c\u67fb\u65b9\u6cd5\u306fPCR\u3092\u5229\u7528\u3057\u305f\u7c21\u4fbf\u3067\u4fe1\u983c\u6027\u306e\u9ad8\u3044\u907a\u4f1d\u5b50\u591a\u578b\u89e3\u6790\u65b9\u6cd5\u306e\u4e00\u7a2e\u3067\u3059\u3002<br \/>\n\u5f53\u5ba4\u304c\u4fdd\u6709\u3057\u3066\u3044\u308b\u5168\u30d2\u30c8\u7531\u6765\u7d30\u80de\u682a\uff08\u5f53\u5ba4\u3067\u57f9\u990a\u3092\u884c\u308f\u306a\u3044\u3082\u306e\u3092\u9664\u304f\uff09\u306b\u95a2\u3057\u3066STR\u691c\u67fb\u3092\u5b9f\u65bd\u3057\u3066\u63d0\u4f9b\u3057\u3066\u3044\u307e\u3059\u3002<\/p>\n<p><strong>1. \u6e96\u5099\u3059\u308b\u3082\u306e<\/strong><\/p>\n<p>(1) \u691c\u4f53<\/p>\n<ul>\n<li>\u691c\u67fb\u7d30\u80de\u682a\u304b\u3089\u62bd\u51fa\u3057\u305f\u30b2\u30ce\u30e0DNA\n<\/ul>\n<p>(2) \u6a5f\u5668\u30fbPCR\u30c1\u30e5\u30fc\u30d6<\/p>\n<ul>\n<li>PCR\u30c1\u30e5\u30fc\u30d6\uff08 0.2ml PCR\u30c1\u30e5\u30fc\u30d6\u3001PCR\u752896\u30a6\u30a7\u30eb\u30d7\u30ec\u30fc\u30c8\u7b49\uff09\n<li>\u30c8\u30ec\u30fc\/\u30ea\u30c6\u30fc\u30ca\u30fc\uff08PCR\u5bb9\u5668\u306b\u9069\u5408\u3059\u308b\u3082\u306e\uff09\n<li>\u30b5\u30fc\u30de\u30eb\u30b5\u30a4\u30af\u30e9\u30fc\uff08Gold 96-well GeneAmp PCR system 9700\u3001Applied Biosystems\uff09\n<li>96well\u6cf3\u52d5\u30d7\u30ec\u30fc\u30c8MicroAmp Optical 96-well Reaction Plate (Applied Biosystems Cat#N8010560 10\u679a)\n<li>Applied Biosystems 3130 Genetic Analyzer\uff08Applied Biosystems\uff09\n<li>3100\/3130\/3730 96-well\u30d7\u30ec\u30fc\u30c8\u30bb\u30d7\u30bf\uff08Applied Biosystems Cat#4315933 20\u679a)\n<li>3130 36-cm Capillary Array\u300047cm\u00d750\u03bcm 4\u672c\/\u30a2\u30ec\u30a4\uff08Applied Biosystems Cat#4333464\uff09\n<li>3100\/3130 96-well \u30d7\u30ec\u30fc\u30c8\u30ea\u30c6\u30fc\u30ca\u30fc\uff08Applied Biosystems Cat#4317241 4\u679a\uff09\n<\/ul>\n<p>(3) PCR Primer<\/p>\n<ul>\n<li>PowerPlex 16 HS System (Promega Cat#DC2101)<br \/>\nKrenke, Benjamin E., et al. &#8220;Validation of a 16-locus fluorescent multiplex system.&#8221; <em>Journal of forensic sciences<\/em> 47.4 (2002): 773-785.\n<\/ul>\n<p>(4) \u8a66\u85ac<\/p>\n<ul>\n<li>PowerPlex 16 HS System (Promega Cat#DC2101)<br \/>\n\u3000\u3000\u30ad\u30c3\u30c8\u5185\u5bb9\uff08\u4f7f\u7528\u3059\u308b\u8a66\u85ac\u306e\u307f\u629c\u7c8b\uff09<br \/>\n\u3000\u3000\u3000PowerPlex HS 5\u00d7Master Mix<br \/>\n\u3000\u3000\u3000PowerPlex 16 HS 10\u00d7Primer Pair Mix<br \/>\n\u3000\u3000\u3000PowerPlex 16 HS Allelic Ladder Mix<br \/>\n\u3000\u3000\u3000Internal Lane Standard(ILS)600<\/p>\n<li>Hi-Di formamide\uff08Applied Biosystems Cat#4311320 25ml\uff09\n<li>POP-4 \u30dd\u30ea\u30de\u30fc\uff08Applied Biosystems Cat#4352755 7.0ml\uff09\n<li>Genetic Analyzer Buffer with EDTA\uff08Applied Biosystems Cat#402824 25ml\uff09\n<li>DDW\n<\/ul>\n<p><strong>2. \u65b9\u6cd5<\/strong><\/p>\n<p>(1) 1st-step PCR<br \/>\n\u3000\u30001)PCR\u30de\u30b9\u30bf\u30fc\u30df\u30c3\u30af\u30b9\u3092\u8abf\u88fd\u3059\u308b\u3002<\/p>\n<table style=\"margin-left:40px;\">\n<tbody>\n<tr>\n<td id=\"t1\" colspan=\"2\" style=\"text-align: right;\">1\u30b5\u30f3\u30d7\u30eb\u3042\u305f\u308a\u306e\u6db2\u91cf<\/td>\n<\/tr>\n<tr>\n<td id=\"t1\">DDW<\/td>\n<td id=\"t1\" style=\"text-align: right; width: 200px;\">17.0\u03bcl<\/td>\n<\/tr>\n<tr>\n<td id=\"t1\">PowerPlex HS 5\u00d7Master Mix<\/td>\n<td id=\"t1\" style=\"text-align: right;\">5.0\u03bcl<\/td>\n<\/tr>\n<tr>\n<td id=\"t1\">PowerPlex 16 HS 10\u00d7Primer Pair Mix<\/td>\n<td id=\"t1\" style=\"text-align: right;\">2.5\u03bcl<\/td>\n<\/tr>\n<tr>\n<td colspan=\"2\" id=\"t1\">\n<hr \/>\n<\/td>\n<\/tr>\n<tr>\n<td id=\"t1\">\u30de\u30b9\u30bf\u30fc\u30df\u30c3\u30af\u30b9\u306e\u6db2\u91cf<\/td>\n<td id=\"t1\" style=\"text-align: right;\">24.5\u03bcl<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<p>\u3000\u30002)PCR\u30c1\u30e5\u30fc\u30d6\u306b\u30de\u30b9\u30bf\u30fc\u30df\u30c3\u30af\u30b9\u309224.5\u03bcl\u305a\u3064\u5206\u6ce8\u3059\u308b\u3002<br \/>\n\u3000\u30003)\u7d041ng\/\u03bcl\u306b\u5e0c\u91c8\u3057\u305f\u30c6\u30f3\u30d7\u30ec\u30fc\u30c8DNA\u30920.5\u03bcl\u305a\u3064\u52a0\u3048\u308b\u3002<br \/>\n\u3000\u30004)PCR\u30c1\u30e5\u30fc\u30d6\u3092\u30c8\u30ec\u30fc\/\u30ea\u30c6\u30fc\u30ca\u30fc\u3068\u5171\u306b\u30b5\u30fc\u30de\u30eb\u30b5\u30a4\u30af\u30e9\u30fc\u306b\u30bb\u30c3\u30c8\u3057\u3001\u4e0b\u8a18\u306e\u30b5\u30a4\u30af\u30eb\u3067\u53cd\u5fdc\u3055\u305b\u308b\u3002<\/p>\n<table style=\"margin-left:40px;\">\n<tbody>\n<tr>\n<td  colspan=\"5\" id=\"t1\">Ramp Speed : 9600<\/td>\n<\/tr>\n<tr>\n<td id=\"t1\" style=\"height: 10px;\"><\/td>\n<\/tr>\n<tr>\n<td id=\"t1\">95\u2103<\/td>\n<td colspan=\"4\" id=\"t1\">11min<\/td>\n<\/tr>\n<tr>\n<td id=\"t1\">96\u2103<\/td>\n<td colspan=\"4\" id=\"t1\">1min<\/td>\n<\/tr>\n<tr>\n<td id=\"t1\" style=\"height: 10px;\"><\/td>\n<\/tr>\n<tr>\n<td id=\"t1\">ramp<\/td>\n<td id=\"t1\">100%<\/td>\n<td id=\"t1\">94\u2103<\/td>\n<td id=\"t1\">30sec<\/td>\n<td id=\"t1\" rowspan=\"3\">for 10 cycle<\/td>\n<\/tr>\n<tr>\n<td id=\"t1\">ramp<\/td>\n<td id=\"t1\" style=\"text-align: center;\">29%<\/td>\n<td id=\"t1\">60\u2103<\/td>\n<td id=\"t1\">30sec<\/td>\n<\/tr>\n<tr>\n<td id=\"t1\">ramp<\/td>\n<td id=\"t1\" style=\"text-align: center;\">23%<\/td>\n<td id=\"t1\">70\u2103<\/td>\n<td id=\"t1\">45sec<\/td>\n<\/tr>\n<tr>\n<td id=\"t1\" style=\"height: 10px;\"><\/td>\n<\/tr>\n<tr>\n<td id=\"t1\">ramp<\/td>\n<td id=\"t1\" style=\"text-align: center;\">100%<\/td>\n<td id=\"t1\">90\u2103<\/td>\n<td id=\"t1\">30sec<\/td>\n<td  id=\"t1\" rowspan=\"3\">for 20 cycle<\/td>\n<\/tr>\n<tr>\n<td id=\"t1\">ramp<\/td>\n<td id=\"t1\" style=\"text-align: center;\">29%<\/td>\n<td id=\"t1\">60\u2103<\/td>\n<td id=\"t1\">30sec<\/td>\n<\/tr>\n<tr>\n<td id=\"t1\">ramp<\/td>\n<td id=\"t1\" style=\"text-align: center;\">23%<\/td>\n<td id=\"t1\">70\u2103<\/td>\n<td id=\"t1\">45sec<\/td>\n<\/tr>\n<tr>\n<td id=\"t1\" style=\"height: 10px;\"><\/td>\n<\/tr>\n<tr>\n<td id=\"t1\">60\u2103<\/td>\n<td  id=\"t1\" colspan=\"4\">30min<\/td>\n<\/tr>\n<tr>\n<td id=\"t1\">4\u2103<\/td>\n<td id=\"t1\" colspan=\"4\">store<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<p>(2) \u5897\u5e45\u65ad\u7247\u306e\u30ad\u30e3\u30d4\u30e9\u30ea\u30fc\u96fb\u6c17\u6cf3\u52d5<\/p>\n<p>\u3000\u30001)\u6cf3\u52d5\u30ab\u30af\u30c6\u30eb\u3092\u8abf\u88fd\u3059\u308b\u3002<br \/>\n\u3000\u3000\u3000\u6cf3\u52d5\u30b5\u30f3\u30d7\u30eb\u6570\u306f\u3001\u691c\u4f53\u6570\uff0b\u30cd\u30ac\u30c6\u30a3\u30d6\u30b3\u30f3\u30c8\u30ed\u30fc\u30eb2\u30b5\u30f3\u30d7\u30eb\uff0bLadder\u30b5\u30f3\u30d7\u30eb\u6570\u3067\u3042\u308b\u3002<\/p>\n<table style=\"margin-left:40px;\">\n<tbody>\n<tr>\n<td id=\"t1\" colspan=\"2\" style=\"text-align: right;\">1\u30b5\u30f3\u30d7\u30eb\u3042\u305f\u308a\u306e\u6db2\u91cf<\/td>\n<\/tr>\n<tr>\n<td id=\"t1\">\u8131\u30a4\u30aa\u30f3\u5316\u30db\u30eb\u30e0\u30a2\u30df\u30c9<\/td>\n<td id=\"t1\" style=\"text-align: right; width: 200px;\">9.75\u03bcl<\/td>\n<\/tr>\n<tr>\n<td id=\"t1\">Internal Lane Standard(ILS)600<\/td>\n<td id=\"t1\" style=\"text-align: right;\">0.25\u03bcl<\/td>\n<\/tr>\n<tr>\n<td colspan=\"2\" id=\"t1\">\n<hr \/>\n<\/td>\n<\/tr>\n<tr>\n<td id=\"t1\">\u30ab\u30af\u30c6\u30eb\u306e\u6db2\u91cf<\/td>\n<td id=\"t1\" style=\"text-align: right;\">10.00\u03bcl<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<p>\u3000\u30002)\u6cf3\u52d5\u30ab\u30af\u30c6\u30eb\u309210\uff5e15\u79d2\u9593\u30df\u30ad\u30b5\u30fc\u3067\u652a\u62cc\u3057\u3001\u6cf3\u52d5\u7528\u30d7\u30ec\u30fc\u30c8\u306b10\u03bcl\u305a\u3064\u5206\u6ce8\u3059\u308b\u3002<br \/>\n\u3000\u30003)\u5897\u5e45\u30b5\u30f3\u30d7\u30eb\u53c8\u306fPowerPlex 16 HS Allelic Ladder Mix\u30921\u03bcl\u305a\u3064\u52a0\u3048\u308b\u3002<br \/>\n\u3000\u30004)\u30bb\u30d7\u30bf\u30b9\u30c8\u30ea\u30c3\u30d7\u3067\u84cb\u3092\u3057\u300195\u2103\u30013\u5206\u9593\u52a0\u71b1\u5f8c\u3001\u76f4\u3061\u306b\u6c37\u4e0a\u306b\u79fb\u30573\u5206\u9593\u51b7\u5374\u3059\u308b\u3002<br \/>\n\u3000\u30005)\u30b5\u30f3\u30d7\u30eb\u30d7\u30ec\u30fc\u30c8\u306e\u4e0a\u306b\u30d7\u30ec\u30fc\u30c8\u30ea\u30c6\u30fc\u30ca\u30fc\u3092\u306f\u3081\u308b\u3002<br \/>\n\u3000\u30006)\u88c5\u7f6e\uff083130 Genetic Analyzer\uff09\u306b\u304b\u3051\u6cf3\u52d5\u3092\u884c\u3046\u3002<\/p>\n<p>(3) \u30c7\u30fc\u30bf\u53ca\u3073\u89e3\u6790<br \/>\nGeneMapper ID\u30bd\u30d5\u30c8\u30a6\u30a7\u30a2\uff08Applied Biosystems\uff09\u306b\u3066\u89e3\u6790\u3092\u884c\u306a\u3046\u3002<\/p>\n<p>a) PowerPlex&#174; 16 HS Allelic Ladder\uff1aPowerPlex 16 HS System\u4ed8\u5c5e\u306e\u30a2\u30ec\u30ea\u30c3\u30af\u30e9\u30c0\u30fc\u30de\u30fc\u30ab\u30fc<br \/>\n<a href=\"\/ja\/wp-content\/uploads\/images\/str_a.jpg\"><img decoding=\"async\" src=\"\/ja\/wp-content\/uploads\/images\/str_a.jpg\" width=\"582px\"><\/a><\/p>\n<p>b) RCB0007 HeLa Lot 46\u306e\u7d50\u679c<br \/>\n\u540c\u3058\u30ed\u30fc\u30ab\u30b9\u306e\u30a2\u30ec\u30ea\u30c3\u30af\u30e9\u30c0\u30fc\u3068\u6a5f\u5668\u306b\u3088\u308a\u6bd4\u8f03\u3059\u308b\u3053\u3068\u3067\u3001\u30a2\u30ec\u30eb\u306e\u5272\u308a\u5f53\u3066\u304c\u884c\u306a\u308f\u308c\u308b\u3002<br \/>\n<a href=\"\/ja\/wp-content\/uploads\/images\/str_b.jpg\"><img decoding=\"async\" src=\"\/ja\/wp-content\/uploads\/images\/str_b.jpg\" width=\"650px\"><\/a><\/p>\n<p>(4) \u5224\u5b9a<br \/>\n\u30c7\u30fc\u30bf\u30d9\u30fc\u30b9\u306e\u30c7\u30fc\u30bf\u3068\u7167\u5408\u3001\u78ba\u8a8d\u3092\u3059\u308b\u3002<br \/>\n\u7d30\u80de\u8a8d\u8a3c\u30c7\u30fc\u30bf\u30d9\u30fc\u30b9<a href=\"http:\/\/jcrbcelldata.nibiohn.go.jp\/str\/\">http:\/\/jcrbcelldata.nibiohn.go.jp\/str\/<\/a><\/p>\n<p>\u3000\u30001)\u7d30\u80de\u682a\u306e\u5224\u5b9a<\/p>\n<p>\u3000\u3000\u3000\u8b58\u5225\u540c\u5b9a\u306b\u3088\u308a\u5408\u683c<\/p>\n<ul>\n<li style=\"margin-left:25px;\">\u767b\u9332\u6e08\u307f\u306e\u4ed6\u306e\u7d30\u80de\u682a\u306e\u4e2d\u306b\u4e00\u81f4\u3059\u308b\u3082\u306e\u304c\u306a\u3044\u2192\u56fa\u6709\u306e\u7d30\u80de\u3000\n<li style=\"margin-left:25px;\">\u767b\u9332\u6e08\u307f\u306e\u4ed6\u306e\u7d30\u80de\u682a\u306e\u4e2d\u306b\u4e00\u81f4\u3059\u308b\u3082\u306e\u304c\u3042\u3063\u3066\u3082\u3001\u6587\u732e\u4e0a\u5225\u540d\u3067\u8a8d\u77e5\u3055\u308c\u3066\u3044\u308b\u7d30\u80de\u306e\u5834\u5408<br \/>\u2192OK\u3068\u306a\u308b\u5834\u5408\u3082\u3042\u308b\n<\/ul>\n<p>\u3000\u3000\u3000\u8b58\u5225\u540c\u5b9a\u306b\u3088\u308a\u4e0d\u5408\u683c<\/p>\n<ul>\n<li style=\"margin-left:25px;\">\u767b\u9332\u6e08\u307f\u306e\u4ed6\u306e\u7d30\u80de\u682a\u306e\u4e2d\u306b\u4e00\u81f4\u3059\u308b\u3082\u306e\u304c\u3042\u308b\u2192\u53d6\u308a\u9055\u3044\n<li style=\"margin-left:25px;\">\u30d7\u30ed\u30c3\u30c8\u30c7\u30fc\u30bf\u4e2d\u306b\u767b\u9332\u6e08\u307f\u306e\u4ed6\u306e\u7d30\u80de\u682a\u306e\u30c7\u30fc\u30bf\u3068\u4e00\u81f4\u3059\u308b\u3082\u306e\u304c\u3042\u308b\u2192\u4ed6\u306e\u7d30\u80de\u6df7\u5165\u306e\u7591\u3044\n<\/ul>\n<p>\u3000\u30002)\u226a \u4ed6\u306e\u7d30\u80de\u306e\u6df7\u5165\u4f8b\u226b\u3000\u30b5\u30f3\u30d7\u30eb\u7d30\u80deC\u306f\u3001\u7d30\u80deA\u3068\u7d30\u80deB\u304c\u6df7\u3056\u3063\u3066\u3044\u308b\u3002<br \/>\n<a href=\"\/ja\/wp-content\/uploads\/images\/str_2.jpg\"><img decoding=\"async\" src=\"\/ja\/wp-content\/uploads\/images\/str_2.jpg\" width=\"653px\"><\/a><\/p>\n","protected":false},"excerpt":{"rendered":"<p>\u30d2\u30c8\u7d30\u80de\u6750\u6599\u306e\u7531\u6765\u500b\u4f53\u306e\u8b58\u5225\u540c\u5b9a\u3001\u8d77\u6e90\u306e\u78ba\u8a8d\u3001\u30af\u30ed\u30b9\u30b3\u30f3\u30bf\u30df\u30cd\u30fc\u30b7\u30e7\u30f3\u306a\u3069\u3092\u78ba\u8a8d\u3059\u308b\u305f\u3081\u306b\u3001Short Tandem Repeat (STR) \u591a\u578b\u89e3\u6790\u3092\u884c\u3063\u3066\u3044\u307e\u3059\u3002\u3053\u306e\u691c\u67fb\u65b9\u6cd5\u306fPCR\u3092\u5229\u7528\u3057\u305f\u7c21\u4fbf\u3067\u4fe1\u983c\u6027\u306e\u9ad8\u3044\u907a\u4f1d\u5b50\u591a\u578b\u89e3\u6790\u65b9\u6cd5\u306e\u4e00\u7a2e\u3067\u3059\u3002 \u5f53\u5ba4\u304c\u4fdd\u6709\u3057\u3066\u3044\u308b\u5168\u30d2\u30c8\u7531\u6765\u7d30\u80de\u682a\uff08\u5f53\u5ba4\u3067\u57f9\u990a\u3092\u884c\u308f\u306a\u3044\u3082\u306e\u3092\u9664\u304f\uff09\u306b\u95a2\u3057\u3066STR\u691c\u67fb\u3092\u5b9f\u65bd\u3057\u3066\u63d0\u4f9b\u3057\u3066\u3044\u307e\u3059\u3002 1. \u6e96\u5099\u3059\u308b\u3082\u306e (1) \u691c\u4f53 \u691c\u67fb 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