{"id":9535,"date":"2016-06-15T10:34:39","date_gmt":"2016-06-15T01:34:39","guid":{"rendered":"http:\/\/cell.brc.riken.jp\/ja\/?page_id=9535"},"modified":"2023-08-10T15:48:07","modified_gmt":"2023-08-10T06:48:07","slug":"sslp","status":"publish","type":"page","link":"http:\/\/cell.brc.riken.jp\/ja\/quality\/sslp","title":{"rendered":"SSLP"},"content":{"rendered":"<p>\u30de\u30a6\u30b9\u7531\u6765\u7d30\u80de\u682a\u306b\u3064\u3044\u3066\u30de\u30a6\u30b9\u7cfb\u7d71\u3092\u540c\u5b9a\u3059\u308b\u691c\u67fb\u3067\u3001PCR\u3092\u5229\u7528\u3057\u305f\u7c21\u4fbf\u3067\u4fe1\u983c\u6027\u306e\u9ad8\u3044\u907a\u4f1d\u5b50\u591a\u578b\u89e3\u6790\u306e\u4e00\u3064\u3067\u3042\u308bSimple Sequence Length Polymorphism\uff08SSLP\uff09 \u6cd5\u3092\u7528\u3044\u3066\u3044\u307e\u3059\u3002<\/p>\n<p>\u5f53\u5ba4\u306eSSLP\u691c\u67fb\u306f\u3001\u6a19\u6e96\u7684\u306b\u591a\u7528\u3055\u308c\u3066\u3044\u308b\u8fd1\u4ea4\u7cfb\u30de\u30a6\u30b9\uff08C57BL\/6, BALB\/c, C3H, 129, DBA\/2\uff09\u306e\u7cfb\u7d71\u3092\u8b58\u5225\u3059\u308b\u3053\u3068\u3092\u76ee\u7684\u3068\u3057\u3066\u958b\u767a\u3057\u305f\u3082\u306e\u3067\u3042\u308a\u3001\uff16\u7b87\u6240\u306e\u30b5\u30c6\u30e9\u30a4\u30c8\u30de\u30fc\u30ab\u30fc\uff08\u591a\u578b\uff09\u3092\u691c\u67fb\u3057\u3066\u3044\u307e\u3059\u3002\u5bc4\u8a17\u8005\u304c\u767b\u9332\u3057\u305f\u7cfb\u7d71\u3092\u78ba\u8a8d\u3059\u308b\u3053\u3068\u304c\u4e3b\u76ee\u7684\u306e\u691c\u67fb\u3067\u3059\u3002\u30de\u30a6\u30b9\u7cfb\u7d71\u306f\u591a\u6570\u5b58\u5728\u3057\u307e\u3059\u306e\u3067\u3001\u4e0a\u8a18\u306e\uff15\u7cfb\u7d71\u3068\u89e3\u6790\u7d50\u679c\u304c\u4e00\u81f4\u3059\u308b\u30de\u30a6\u30b9\u7cfb\u7d71\u304c\u4ed6\u306b\u3082\u5b58\u5728\u3059\u308b\u53ef\u80fd\u6027\u3092\u5426\u5b9a\u3067\u304d\u307e\u305b\u3093\u3002<br \/>\n\u5f93\u3044\u307e\u3057\u3066\u3001\u5f53\u5ba4\u304c\u5b9f\u65bd\u3057\u3066\u3044\u308bSSLP\u691c\u67fb\u306e\u7d50\u679c\u3092\u3082\u3063\u3066\u3001\u30de\u30a6\u30b9\u7d30\u80de\u306e\u7531\u6765\u7cfb\u7d71\u3092\u4fdd\u8a3c\u3059\u308b\u3082\u306e\u3067\u306f\u306a\u3044\u3053\u3068\u3092\u3054\u4e86\u627f\u304f\u3060\u3055\u3044\u3002<\/p>\n<p><strong>\u53c2\u8003\u6587\u732e<\/strong><br \/>\nTsang S, Sun Z, et al. ; Mamm Genome. 2005 Jul; 16(7):476-80. (PMID: 16151692)<br \/>\nBerg KD, et al. ; J Mol Diagn. 2000 Feb; 2(1):20-8. (PMID: 11272898)<br \/>\nBeck JA, et al. ; Nat Genet. 2000 Jan; 24(1):23-5. (PMID: 10615122)<\/p>\n<p><strong>1. \u6e96\u5099\u3059\u308b\u3082\u306e<\/strong><\/p>\n<p>(1) \u691c\u4f53<\/p>\n<ul>\n<li>\u7d30\u80de\u304b\u3089\u62bd\u51fa\u3057\u305f\u30b2\u30ce\u30e0DNA : 25ng\/\u03bcl\u3092\u30c6\u30f3\u30d7\u30ec\u30fc\u30c8DNA\u3068\u3059\u308b\u3002\n<\/ul>\n<p>(2) \u6a5f\u5668\u30fbPCR\u30d7\u30ec\u30fc\u30c8\u7b49<\/p>\n<ul>\n<li>PCR\u752896well\u30d7\u30ec\u30fc\u30c8\u3001\u30d7\u30ec\u30fc\u30c8\u30ab\u30d0\u30fc\n<li>\u30c8\u30ec\u30fc\/\u30ea\u30c6\u30fc\u30ca\u30fc\uff08PCR\u5bb9\u5668\u306b\u9069\u5408\u3059\u308b\u3082\u306e\uff09\n<li>\u30d7\u30ec\u30fc\u30c8\u30b7\u30fc\u30eb\uff08\u7c98\u7740\u5f0f\uff09\n<li>\u30b5\u30fc\u30de\u30eb\u30b5\u30a4\u30af\u30e9\u30fc\uff08Gold 96-well GeneAmp PCR system 9700\u3001Applied Biosystems\uff09\n<li>\u5e0c\u91c8\u5bb9\u5668\uff08\u30c1\u30e5\u30fc\u30d6\u3084\u30d7\u30ec\u30fc\u30c8\u7b49\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<li>3130 POP-4 \u30dd\u30ea\u30de\u30fc\uff08Applied Biosystems Cat#4352755 7.0ml\uff09\n<\/ul>\n<p>(3) PCR\u30d7\u30e9\u30a4\u30de\u30fc<\/p>\n<ul>\n<li>ABI PRISM Mouse Mapping Primers v1.0 (Applied Biosystems 3000pmol 5\u03bcM each primer)\n<\/ul>\n<table>\n<tbody>\n<th>\u30d7\u30e9\u30a4\u30de\u30fc\u540d\uff08\u7565\u79f0\uff09<\/th>\n<th>\u30de\u30fc\u30ab\u30fc<\/th>\n<th>\u86cd\u5149\u6a19\u8b58<\/th>\n<th>\u30ab\u30bf\u30ed\u30b0\u756a\u53f7<\/th>\n<tr>\n<td>1G<\/td>\n<td>D1Mit159.1<\/td>\n<td>VIC(\u7dd1)<\/td>\n<td>4323000<\/td>\n<\/tr>\n<tr>\n<td>2B<\/td>\n<td>D2Mit395.1<\/td>\n<td>6-FAM(\u9752)<\/td>\n<td>4323128<\/td>\n<\/tr>\n<tr>\n<td>23B<\/td>\n<td>D4Mit170.1<\/td>\n<td>6-FAM(\u9752)<\/td>\n<td>4323227<\/td>\n<\/tr>\n<tr>\n<td>4G<\/td>\n<td>D5Mit201.1<\/td>\n<td>VIC(\u7dd1)<\/td>\n<td>4323237<\/td>\n<\/tr>\n<tr>\n<td>11Y<\/td>\n<td>D13Mit256.1<\/td>\n<td>NED(\u9ec4)<\/td>\n<td>4323745<\/td>\n<\/tr>\n<tr>\n<td>15Y<\/td>\n<td>D17Mit51.1<\/td>\n<td>NED(\u9ec4)<\/td>\n<td>4323803<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<p>\u203b\u8ca9\u58f2\u4e2d\u6b62\u3060\u304c\u3001Primer\u306e\u30ab\u30bf\u30ed\u30b0\u756a\u53f7\u304b\u3089\u4f5c\u6210\u3057\u3066\u3082\u3089\u3048\u308b<\/p>\n<p>(4) \u8a66\u85ac<\/p>\n<ul>\n<li>True Allele PCR Premix (Applied Biosystems Cat#403061 2000\u53cd\u5fdc\u5206)\n<li>GeneScan \u2013500 LIZ Size Standerd (Applied Biosystems Cat#4322682 800\u30ec\u30fc\u30f3\u5206)\u3000\u86cd\u5149\u6a19\u8b58\u306fLIZ\uff08\u6a59\uff09\n<li>Hi-Di formamide\uff08Applied Biosystems Cat#4311320 25ml\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><br \/>\n(1) PCR<br \/>\n\u3000\u30001) \u5404\u30d7\u30e9\u30a4\u30de\u30fc\uff081G\u30012B\u300111Y\u300123B\u300115Y\u306e\u6fc3\u5ea6\u306f\u539f\u6db2\u306e\u307e\u307e5\u03bcM\u30014G\u306e\u307f1\/5\u306b\u5e0c\u91c8\u30571\u03bcM\uff09\u30921\u03bcl\u305a\u3064\u3001PCR\u752896 well\u30d7\u30ec\u30fc\u30c8\u306e\u30a6\u30a7\u30eb\u306b\u5165\u308c\u308b\u3002<br \/>\n\u3000\u30002) True Allele PCR Premix\u5e0c\u91c8\u6db2\u3092\u5404\u30a6\u30a7\u30eb\u306b11.6\u03bcl\u305a\u3064\u52a0\u3048\u308b\u3002<\/p>\n<ul>\n<li>True Allele PCR Premix\u5e0c\u91c8\u6db2\u306e\u7d44\u6210\n<\/ul>\n<table style=\"margin-left:40px;\">\n<tbody>\n<tr>\n<td id=\"t1\">DDW<\/td>\n<td id=\"t1\" style=\"text-align: right;\">2.6\u03bcl<\/td>\n<\/tr>\n<tr>\n<td id=\"t1\">True Allele PCR Premix<\/td>\n<td id=\"t1\" style=\"text-align: right;\">9.0\u03bcl<\/td>\n<\/tr>\n<tr>\n<td colspan=\"2\" id=\"t1\">\n<hr \/>\n<\/td>\n<\/tr>\n<tr>\n<td id=\"t1\">\u5e0c\u91c8\u6db2\u306e\u6db2\u91cf<\/td>\n<td id=\"t1\" style=\"text-align: right;\">11.6\u03bcl<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<p>\u3000\u30003) \u691c\u4f53\u3092DDW\u306725ng\/\u03bcl\u306b\u5e0c\u91c8\u3057\u3001\u5404\u30a6\u30a7\u30eb\u306b2.4\u03bcl\u305a\u3064\u52a0\u3048\u308b\u3002<br \/>\n\u3000\u30004) PCR\u752896 well\u30d7\u30ec\u30fc\u30c8\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\u3059\u308b\u3002<br \/>\n\u3000\u30005) PCR program<br \/>\n\u3000\u3000\u3000Ramp Speed : 9600<br \/>\n\u3000\u3000\u3000Stage1 : 95\u2103\u300012\u5206\uff081\u30b5\u30a4\u30af\u30eb\uff09<br \/>\n\u3000\u3000\u3000Stage2 : 94\u2103  20\u79d2\u3000\u2192\u300055\u2103\u300020\u79d2\u3000\u2192\u300072\u2103\u300030\u79d2\uff0810\u30b5\u30a4\u30af\u30eb\uff09<br \/>\n\u3000\u3000\u3000Stage3 : 89\u2103\u300020\u79d2\u3000\u2192\u300055\u2103\u300020\u79d2\u3000\u2192\u300072\u2103\u300030\u79d2\uff0820\u30b5\u30a4\u30af\u30eb\uff09<br \/>\n\u3000\u3000\u3000Stage4 : 72\u2103\u300010\u5206\uff081\u30b5\u30a4\u30af\u30eb\uff09<br \/>\n\u3000\u3000\u3000Stage5 : 4\u2103\u3000\uff08\u4fdd\u5b58\uff09<\/p>\n<p>(2) \u5897\u5e45\u65ad\u7247\u306e\u30ad\u30e3\u30d4\u30e9\u30ea\u30fc\u96fb\u6c17\u6cf3\u52d5<br \/>\n\u9752\u3001\u9ec4\u3001\u7dd13\u8272\u306e\u5897\u5e45\u30b5\u30f3\u30d7\u30eb\u306f\u30de\u30fc\u30ab\u30fc\u30b0\u30eb\u30fc\u30d7\u3054\u3068\u306b1\u672c\u306b\u307e\u3068\u3081\u3066\u6cf3\u52d5\u3059\u308b\u3002<\/p>\n<ul>\n<li>Group1\u3000\u30001G  2B  11Y\n<li>Group3\u3000\u30004G  15Y  23B\n<\/ul>\n<p>\u3000\u30001) \u5897\u5e45\u30b5\u30f3\u30d7\u30eb\u30922\u03bcl\u305a\u3064\u5e0c\u91c8\u5bb9\u5668\u306b\u79fb\u3059\u3002\u3053\u306e\u6642\u30b0\u30eb\u30fc\u30d7\u3054\u3068\u306b3\u30a6\u30a7\u30eb\u306e\u30b5\u30f3\u30d7\u30eb\u30921\u672c\uff08\u53c8\u306f1\u30a6\u30a7\u30eb\uff09\u306b\u307e\u3068\u3081\u308b\u3002<br \/>\n\u3000\u30002) DDW\u309214\u03bcl\u305a\u3064\u52a0\u3048\u3001\u30d4\u30da\u30c3\u30c6\u30a3\u30f3\u30b0\u306b\u3088\u308a\u6df7\u548c\u3059\u308b\u3002\uff08\u5404\u5897\u5e45\u30b5\u30f3\u30d7\u30eb\u304c1\/10\u5e0c\u91c8\u306b\u306a\u308b\u3002\uff09<br \/>\n\u3000\u30003) \u6cf3\u52d5\u30ab\u30af\u30c6\u30eb\u3092\u8abf\u88fd\u3059\u308b\u3002<br \/>\n\u3000\u3000\u3000\u203b\u3000\u6cf3\u52d5\u30b5\u30f3\u30d7\u30eb\u6570\u306f1\u691c\u4f53\u306b\u3064\u304d2\u30b5\u30f3\u30d7\u30eb\u3002<\/p>\n<ul>\n<li>\u6cf3\u52d5\u30ab\u30af\u30c6\u30eb\u306e\u7d44\u6210\n<\/ul>\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;\">19.5\u03bcl<\/td>\n<\/tr>\n<tr>\n<td id=\"t1\">Size Standard<\/td>\n<td id=\"t1\" style=\"text-align: right;\">0.5\u03bcl<\/td>\n<\/tr>\n<tr>\n<td colspan=\"2\" id=\"t1\">\n<hr \/>\n<\/td>\n<\/tr>\n<tr>\n<td id=\"t1\">\u30ab\u30af\u30c6\u30eb\u306e\u6db2\u91cf<\/td>\n<td id=\"t1\" style=\"text-align: right;\">20.0\u03bcl<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<p>\u3000\u30004) \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\u306b20\u03bcl\u305a\u3064\u5206\u6ce8\u3059\u308b\u3002<br \/>\n\u3000\u30005) \u5897\u5e45\u30b5\u30f3\u30d7\u30eb\u5e0c\u91c8\u6db2\u30921\u03bcl\u305a\u3064\u52a0\u3048\u308b\u3002<br \/>\n\u3000\u30006) \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\u30007) \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\u30008) \u88c5\u7f6e\uff083130 Genetic Analyzer\uff09\u306b\u304b\u3051\u6cf3\u52d5\u3092\u884c\u3046\u3002<\/p>\n<p>(3) \u89e3\u6790<br \/>\nABI PRISM GeneMapper v4.0\u306b\u3066\u89e3\u6790\u3092\u884c\u3046\u3002<\/p>\n<p>\u89e3\u6790\u4f8b<br \/>\n<img decoding=\"async\" src=\"\/ja\/wp-content\/uploads\/images\/sslp_ex.jpg\"><\/a><\/p>\n","protected":false},"excerpt":{"rendered":"<p>\u30de\u30a6\u30b9\u7531\u6765\u7d30\u80de\u682a\u306b\u3064\u3044\u3066\u30de\u30a6\u30b9\u7cfb\u7d71\u3092\u540c\u5b9a\u3059\u308b\u691c\u67fb\u3067\u3001PCR\u3092\u5229\u7528\u3057\u305f\u7c21\u4fbf\u3067\u4fe1\u983c\u6027\u306e\u9ad8\u3044\u907a\u4f1d\u5b50\u591a\u578b\u89e3\u6790\u306e\u4e00\u3064\u3067\u3042\u308bSimple Sequence Length Polymorphism\uff08SSLP\uff09 \u6cd5\u3092\u7528\u3044\u3066\u3044\u307e\u3059\u3002 \u5f53\u5ba4\u306eSSLP\u691c\u67fb\u306f\u3001\u6a19\u6e96\u7684\u306b\u591a\u7528\u3055\u308c\u3066\u3044\u308b\u8fd1\u4ea4\u7cfb\u30de\u30a6\u30b9\uff08C57BL\/6, BALB\/c, C3H, 129, DBA\/2\uff09\u306e\u7cfb\u7d71\u3092\u8b58\u5225\u3059\u308b\u3053\u3068\u3092\u76ee\u7684\u3068\u3057\u3066\u958b\u767a\u3057\u305f\u3082\u306e\u3067\u3042\u308a\u3001\uff16\u7b87\u6240\u306e [&hellip;]<\/p>\n","protected":false},"author":14,"featured_media":0,"parent":3425,"menu_order":0,"comment_status":"closed","ping_status":"closed","template":"","meta":{"_seopress_robots_primary_cat":"","_seopress_titles_title":"","_seopress_titles_desc":"","_seopress_robots_index":"","_mc_calendar":[],"footnotes":"","_wp_rev_ctl_limit":""},"class_list":["post-9535","page","type-page","status-publish","hentry"],"_links":{"self":[{"href":"http:\/\/cell.brc.riken.jp\/ja\/wp-json\/wp\/v2\/pages\/9535","targetHints":{"allow":["GET"]}}],"collection":[{"href":"http:\/\/cell.brc.riken.jp\/ja\/wp-json\/wp\/v2\/pages"}],"about":[{"href":"http:\/\/cell.brc.riken.jp\/ja\/wp-json\/wp\/v2\/types\/page"}],"author":[{"embeddable":true,"href":"http:\/\/cell.brc.riken.jp\/ja\/wp-json\/wp\/v2\/users\/14"}],"replies":[{"embeddable":true,"href":"http:\/\/cell.brc.riken.jp\/ja\/wp-json\/wp\/v2\/comments?post=9535"}],"version-history":[{"count":6,"href":"http:\/\/cell.brc.riken.jp\/ja\/wp-json\/wp\/v2\/pages\/9535\/revisions"}],"predecessor-version":[{"id":13819,"href":"http:\/\/cell.brc.riken.jp\/ja\/wp-json\/wp\/v2\/pages\/9535\/revisions\/13819"}],"up":[{"embeddable":true,"href":"http:\/\/cell.brc.riken.jp\/ja\/wp-json\/wp\/v2\/pages\/3425"}],"wp:attachment":[{"href":"http:\/\/cell.brc.riken.jp\/ja\/wp-json\/wp\/v2\/media?parent=9535"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}