{"id":10110,"date":"2017-01-17T15:13:52","date_gmt":"2017-01-17T06:13:52","guid":{"rendered":"http:\/\/cell.brc.riken.jp\/ja\/?page_id=10110"},"modified":"2018-02-01T17:47:29","modified_gmt":"2018-02-01T08:47:29","slug":"pcr","status":"publish","type":"page","link":"http:\/\/cell.brc.riken.jp\/ja\/quality\/pcr","title":{"rendered":"\u52d5\u7269\u7a2e\u3092\u540c\u5b9a\u3059\u308bPCR\u691c\u67fb"},"content":{"rendered":"<p>\u7d30\u80de\u682a\u306e\u7531\u6765\u52d5\u7269\u7a2e\u306e\u540c\u5b9a\u3068\u5224\u5225\u53ef\u80fd\u306a\u52d5\u7269\u7a2e\uff08\u6ce81\uff09\u306e\u6df7\u5165\u306a\u3069\u306e\u30af\u30ed\u30b9\u30b3\u30f3\u30bf\u30df\u30cd\u30fc\u30b7\u30e7\u30f3\u306e\u6709\u7121\u3092\u78ba\u8a8d\u3059\u308b\u305f\u3081\u3001\u52d5\u7269\u7a2e\u3092\u540c\u5b9a\u3059\u308bPCR\u691c\u67fb\u3092\u3057\u3066\u3044\u307e\u3059\u3002\u73fe\u5728\u3001\u7d30\u80de\u57f9\u990a\u306b\u5e83\u7bc4\u306b\u7528\u3044\u3089\u308c\u308b\u52d5\u7269\u7a2e13\u7a2e\u306b\u3064\u3044\u3066\u52d5\u7269\u7a2e\u306e\u5224\u5b9a\u304c\u53ef\u80fd\u3067\u3059\u3002<br \/>\n\u6ce81\uff09\u5224\u5225\u53ef\u80fd\u306a\u52d5\u7269\u7a2e<br \/>\n\u30d2\u30c8\u3001\u30de\u30a6\u30b9\u3001\u30e9\u30c3\u30c8\u3001\u30cb\u30ef\u30c8\u30ea\u3001\u30b7\u30ea\u30a2\u30f3\u30cf\u30e0\u30b9\u30bf\u30fc\u3001\u30a2\u30d5\u30ea\u30ab\u30df\u30c9\u30ea\u30b6\u30eb\u3001\u30ab\u30cb\u30af\u30a4\u30b6\u30eb\u3001\u30a6\u30b5\u30ae\u3001\u30c1\u30e3\u30a4\u30cb\u30fc\u30ba\u30cf\u30e0\u30b9\u30bf\u30fc\u3001\u30a4\u30cc\u3001\u30d6\u30bf\u3001\u30a6\u30b7\u3001\u30cd\u30b3<\/p>\n<p>\u5185\u90e8\u30b3\u30f3\u30c8\u30ed\u30fc\u30eb\u30d7\u30e9\u30a4\u30de\u30fc ( 18SrRNA ) \u3068\u52d5\u7269\u7a2e\u7279\u7570\u7684\u30d7\u30e9\u30a4\u30de\u30fc\u3092\u7528\u3044\u3066\u3001\u7d30\u80de\u4e2d\u306b\u542b\u307e\u308c\u308b\u30df\u30c8\u30b3\u30f3\u30c9\u30ea\u30a2DNA \u306e\u5404\u52d5\u7269\u7279\u7570\u7684\u914d\u5217\u3092PCR\u6cd5\u3067\u691c\u51fa\u3059\u308b\u3053\u3068\u306b\u3088\u308a\u3001\u52d5\u7269\u7a2e\u3092\u540c\u5b9a\u3057\u3066\u3044\u307e\u3059\u3002\u77ed\u6642\u9593\u3067\u540c\u5b9a\u3059\u308b\u3053\u3068\u304c\u3067\u304d\u307e\u3059\u304c\u3001\u5fae\u91cf\u306e\u6df7\u5165\u306f\u691c\u51fa\u9650\u754c\u304c\u3042\u308b\u305f\u3081\u691c\u51fa\u3055\u308c\u306a\u3044\u3053\u3068\u3082\u3042\u308a\u307e\u3059\u3002<\/p>\n<p>\uff1c\u53c2\u8003\u6587\u732e\uff1e<br \/>\n<a href=\"https:\/\/link.springer.com\/article\/10.1007%2Fs11626-007-9033-5\">In Vitro Cell.Dev.Biol.\u2014Animal (2007) 43:168\u2013175 <\/a><\/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<\/li>\n<li>\u9670\u6027\u5bfe\u7167\uff1aPCR\u30de\u30b9\u30bf\u30fc\u30df\u30c3\u30af\u30b9\u4f5c\u6210\u6642\u306b\u4f7f\u7528\u3057\u305f\u84b8\u7559\u7cbe\u88fd\u6c34<\/li>\n<li>\u30af\u30ed\u30b9\u30b3\u30f3\u30bf\u30df\u30cd\u30fc\u30b7\u30e7\u30f3\u78ba\u8a8d\u306e\u305f\u3081\u306e\u967d\u6027\u5bfe\u7167\uff08\u4e00\u822c\u306b\u53d6\u308a\u6271\u3044\u306e\u591a\u3044\u52d5\u7269\u7a2e\uff09\n<ul>\n<li>\u967d\u6027\u5bfe\u71671\uff1a\u30d2\u30c8\u306eDNA<\/li>\n<li>\u967d\u6027\u5bfe\u71671\uff1a\u30de\u30a6\u30b9\u306eDNA<\/li>\n<li>\u967d\u6027\u5bfe\u71671\uff1a\u30e9\u30c3\u30c8\u306eDNA<\/li>\n<\/ul>\n<\/li>\n<li>\u691c\u67fb\u7d30\u80de\u682a\u306e\u52d5\u7269\u7a2e\u540c\u5b9a\u306e\u305f\u3081\u306e\u967d\u6027\u5bfe\u7167\uff08\u967d\u6027\u5bfe\u71671\u4ee5\u5916\u306e\u52d5\u7269\u7a2e\u306e\u5834\u5408\uff09<\/li>\n<li>\u967d\u6027\u5bfe\u71672\uff1a\u691c\u67fb\u7d30\u80de\u682a\u3068\u540c\u3058\u52d5\u7269\u7a2e\u306eDNA<br \/>\n\u30d2\u30c8\u3001\u30de\u30a6\u30b9\u3001\u30e9\u30c3\u30c8\u306e\u5834\u5408\u306f\u967d\u6027\u5bfe\u7167\uff11\u3067\u517c\u306d\u308b\u3002<\/li>\n<\/ul>\n<p>(2)\u6a5f\u5668\u3068PCR\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<\/li>\n<li>\u30c8\u30ec\u30fc\/\u30ea\u30c6\u30fc\u30ca\u30fc\uff08 PCR\u5bb9\u5668\u306b\u9069\u5408\u3059\u308b\u3082\u306e \uff09<\/li>\n<li>\u30b5\u30fc\u30de\u30eb\u30b5\u30a4\u30af\u30e9\u30fc\uff08 Gold 96-well GeneAmp PCR system 9700 \u3001Applied Biosystems \uff09<\/li>\n<li>\u30a2\u30ac\u30ed\u30fc\u30b9\u30b2\u30eb\u96fb\u6c17\u6cf3\u52d5\u88c5\u7f6e\uff08 Mupid\u30b7\u30ea\u30fc\u30ba\u3000ADVANCE \uff09<\/li>\n<li>\u30b2\u30eb\u30c8\u30ec\u30a4 \u3001\u30b3\u30fc\u30e0\uff08\u3000\u30b2\u30eb\u30c8\u30ec\u30a4-L 25well\u3001\u30b2\u30eb\u30c8\u30ec\u30a4-S 12well\u306e\u30b5\u30a4\u30ba\uff09<\/li>\n<\/ul>\n<p>(3)\u30d7\u30e9\u30a4\u30de\u30fc\u4e00\u89a7<\/p>\n<table border=\"1\">\n<tbody>\n<tr>\n<th style=\"text-align: center;\">\u52d5\u7269\u7a2e<\/th>\n<th style=\"text-align: center;\"><\/th>\n<th style=\"text-align: center;\">\u30d7\u30e9\u30a4\u30de\u30fc\u540d<\/th>\n<th style=\"text-align: center;\">\u914d\u5217<\/th>\n<th style=\"text-align: center;\">\u30b5\u30a4\u30ba(bp)<\/th>\n<th style=\"text-align: center;\">\u53c2\u7167\u914d\u5217<\/th>\n<\/tr>\n<tr>\n<td style=\"text-align: center; background-color: #ffffff;\" rowspan=\"2\">Human<\/td>\n<td style=\"background-color: #ffffff;\">FW<\/td>\n<td style=\"background-color: #ffffff;\">S31_Human_F2<\/td>\n<td style=\"background-color: #ffffff;\">CTCCTATTCTTGCACGAAAC<\/td>\n<td style=\"text-align: center; background-color: #ffffff;\" rowspan=\"2\">330<\/td>\n<td style=\"text-align: center; background-color: #ffffff;\" rowspan=\"2\">NC 001807<\/td>\n<\/tr>\n<tr>\n<td style=\"background-color: #ffffff;\">RV<\/td>\n<td style=\"background-color: #ffffff;\">S53_Human_R7<\/td>\n<td style=\"background-color: #ffffff;\">GATGGGGATTATTGCTAGGATG<\/td>\n<\/tr>\n<tr>\n<td style=\"text-align: center; background-color: #ffffff;\" rowspan=\"2\">Mouse<\/td>\n<td style=\"background-color: #ffffff;\">FW<\/td>\n<td style=\"background-color: #ffffff;\">S07_Mouse_F<\/td>\n<td style=\"background-color: #ffffff;\">GCACTGAAAATGCTTAGATGGATAATTG<\/td>\n<td style=\"text-align: center; background-color: #ffffff;\" rowspan=\"2\">948<\/td>\n<td style=\"text-align: center; background-color: #ffffff;\" rowspan=\"2\">NC 005089<\/td>\n<\/tr>\n<tr>\n<td style=\"background-color: #ffffff;\">RV<\/td>\n<td style=\"background-color: #ffffff;\">S08_Mouse_R<\/td>\n<td style=\"background-color: #ffffff;\">CCTCTCATAAACGGATGTCTAG<\/td>\n<\/tr>\n<tr>\n<td style=\"text-align: center; background-color: #ffffff;\" rowspan=\"2\">Rat<\/td>\n<td style=\"background-color: #ffffff;\">FW<\/td>\n<td style=\"background-color: #ffffff;\">S29_Rat_F2<\/td>\n<td style=\"background-color: #ffffff;\">CAATTCTCCTAGCACAAGTG<\/td>\n<td style=\"text-align: center; background-color: #ffffff;\" rowspan=\"2\">493<\/td>\n<td style=\"text-align: center; background-color: #ffffff;\" rowspan=\"2\">NC 001665.2<\/td>\n<\/tr>\n<tr>\n<td style=\"background-color: #ffffff;\">RV<\/td>\n<td style=\"background-color: #ffffff;\">S30_Rat_R2<\/td>\n<td style=\"background-color: #ffffff;\">CCCCAACCGAAATTTTTTAGTTC<\/td>\n<\/tr>\n<tr>\n<td style=\"text-align: center; background-color: #ffffff;\" rowspan=\"2\">Chicken<\/td>\n<td style=\"background-color: #ffffff;\">FW<\/td>\n<td style=\"background-color: #ffffff;\">S27_Chicken_F<\/td>\n<td style=\"background-color: #ffffff;\">GTATTCCCGTGCAAAAACGAG<\/td>\n<td style=\"text-align: center; background-color: #ffffff;\" rowspan=\"2\">197<\/td>\n<td style=\"text-align: center; background-color: #ffffff;\" rowspan=\"2\">AB086102<\/td>\n<\/tr>\n<tr>\n<td style=\"background-color: #ffffff;\">RV<\/td>\n<td style=\"background-color: #ffffff;\">S28_Chicken_R<\/td>\n<td style=\"background-color: #ffffff;\">CTTAGTGAAGAGTTGTGGTCTG<\/td>\n<\/tr>\n<tr>\n<td style=\"text-align: center; background-color: #ffffff;\" rowspan=\"2\">Syrian hamster<\/td>\n<td style=\"background-color: #ffffff;\">FW<\/td>\n<td style=\"background-color: #ffffff;\">S11_S. hamster_F<\/td>\n<td style=\"background-color: #ffffff;\">GACCTCTTAGGTGTATTCCTAC<\/td>\n<td style=\"text-align: center; background-color: #ffffff;\" rowspan=\"2\">245<\/td>\n<td style=\"text-align: center; background-color: #ffffff;\" rowspan=\"2\">AF119265<\/td>\n<\/tr>\n<tr>\n<td style=\"background-color: #ffffff;\">RV<\/td>\n<td style=\"background-color: #ffffff;\">S12_S. hamster_R<\/td>\n<td style=\"background-color: #ffffff;\">GTATGAAGAAGGGGTAGAGCA<\/td>\n<\/tr>\n<tr>\n<td style=\"text-align: center; background-color: #ffffff;\" rowspan=\"2\">African green monkey<\/td>\n<td style=\"background-color: #ffffff;\">FW<\/td>\n<td style=\"background-color: #ffffff;\">S54_A. monkey_F3<\/td>\n<td style=\"background-color: #ffffff;\">AAATCAAGGCATAGCTTAACGC<\/td>\n<td style=\"text-align: center; background-color: #ffffff;\" rowspan=\"2\">576<\/td>\n<td style=\"text-align: center; background-color: #ffffff;\" rowspan=\"2\">AY863426.1<\/td>\n<\/tr>\n<tr>\n<td style=\"background-color: #ffffff;\">RV<\/td>\n<td style=\"background-color: #ffffff;\">S55_A. monkey_R3<\/td>\n<td style=\"background-color: #ffffff;\">GGCCAACTATGGTAGTTATGGT<\/td>\n<\/tr>\n<tr>\n<td style=\"text-align: center; background-color: #ffffff;\" rowspan=\"2\">Cynomolgus monkey<\/td>\n<td style=\"background-color: #ffffff;\">FW<\/td>\n<td style=\"background-color: #ffffff;\">S43_C. monkey_F3<\/td>\n<td style=\"background-color: #ffffff;\">CCTAAACCTCAGTAGTTAGATA<\/td>\n<td style=\"text-align: center; background-color: #ffffff;\" rowspan=\"2\">577<\/td>\n<td style=\"text-align: center; background-color: #ffffff;\" rowspan=\"2\">AF424970<\/td>\n<\/tr>\n<tr>\n<td style=\"background-color: #ffffff;\">RV<\/td>\n<td style=\"background-color: #ffffff;\">S44_C. monkey_R3<\/td>\n<td style=\"background-color: #ffffff;\">CTCAAGGTAATCAAGTGGTAT<\/td>\n<\/tr>\n<tr>\n<td style=\"text-align: center; background-color: #ffffff;\" rowspan=\"2\">Rabbit<\/td>\n<td style=\"background-color: #ffffff;\">FW<\/td>\n<td style=\"background-color: #ffffff;\">S35_Rabbit_F2<\/td>\n<td style=\"background-color: #ffffff;\">CGGACATTAAGCACACTAATC<\/td>\n<td style=\"text-align: center; background-color: #ffffff;\" rowspan=\"2\">610<\/td>\n<td style=\"text-align: center; background-color: #ffffff;\" rowspan=\"2\">NC 001913<\/td>\n<\/tr>\n<tr>\n<td style=\"background-color: #ffffff;\">RV<\/td>\n<td style=\"background-color: #ffffff;\">S36_Rabbit_R2<\/td>\n<td style=\"background-color: #ffffff;\">CATTGCTCTCCACTCTATGG<\/td>\n<\/tr>\n<tr>\n<td style=\"text-align: center; background-color: #ffffff;\" rowspan=\"2\">Chinese hamster<\/td>\n<td style=\"background-color: #ffffff;\">FW<\/td>\n<td style=\"background-color: #ffffff;\">S13_C. hamster_F<\/td>\n<td style=\"background-color: #ffffff;\">CCGGCGTAAAACGTGTTATAGACT<\/td>\n<td style=\"text-align: center; background-color: #ffffff;\" rowspan=\"2\">601<\/td>\n<td style=\"text-align: center; background-color: #ffffff;\" rowspan=\"2\">DQ390542<\/td>\n<\/tr>\n<tr>\n<td style=\"background-color: #ffffff;\">RV<\/td>\n<td style=\"background-color: #ffffff;\">S14_C. hamster_R<\/td>\n<td style=\"background-color: #ffffff;\">GTATTAGGTATAATATCGGCAGTC<\/td>\n<\/tr>\n<tr>\n<td style=\"text-align: center; background-color: #ffffff;\" rowspan=\"2\">Dog<\/td>\n<td style=\"background-color: #ffffff;\">FW<\/td>\n<td style=\"background-color: #ffffff;\">S19_Dog_F<\/td>\n<td style=\"background-color: #ffffff;\">GCCCAACTAACCCCAAACTTA<\/td>\n<td style=\"text-align: center; background-color: #ffffff;\" rowspan=\"2\">755<\/td>\n<td style=\"text-align: center; background-color: #ffffff;\" rowspan=\"2\">AY729880<\/td>\n<\/tr>\n<tr>\n<td style=\"background-color: #ffffff;\">RV<\/td>\n<td style=\"background-color: #ffffff;\">S20_Dog_R<\/td>\n<td style=\"background-color: #ffffff;\">GGTTAACAATGGGGTGGATAAG<\/td>\n<\/tr>\n<tr>\n<td style=\"text-align: center; background-color: #ffffff;\" rowspan=\"2\">Pig<\/td>\n<td style=\"background-color: #ffffff;\">FW<\/td>\n<td style=\"background-color: #ffffff;\">S25_Pig_F<\/td>\n<td style=\"background-color: #ffffff;\">CCTATATTCAATTACACAACCATGC<\/td>\n<td style=\"text-align: center; background-color: #ffffff;\" rowspan=\"2\">819<\/td>\n<td style=\"text-align: center; background-color: #ffffff;\" rowspan=\"2\">AY337045<\/td>\n<\/tr>\n<tr>\n<td style=\"background-color: #ffffff;\">RV<\/td>\n<td style=\"background-color: #ffffff;\">S26_Pig_R<\/td>\n<td style=\"background-color: #ffffff;\">GCGTGTGCGAGGAGAAAGGC<\/td>\n<\/tr>\n<tr>\n<td style=\"text-align: center; background-color: #ffffff;\" rowspan=\"2\">Cow<\/td>\n<td style=\"background-color: #ffffff;\">FW<\/td>\n<td style=\"background-color: #ffffff;\">S23_Cow_F<\/td>\n<td style=\"background-color: #ffffff;\">CCTAGATGAGTCTCCCAACTC<\/td>\n<td style=\"text-align: center; background-color: #ffffff;\" rowspan=\"2\">1090<\/td>\n<td style=\"text-align: center; background-color: #ffffff;\" rowspan=\"2\">AB074965<\/td>\n<\/tr>\n<tr>\n<td style=\"background-color: #ffffff;\">RV<\/td>\n<td style=\"background-color: #ffffff;\">S24_Cow_R<\/td>\n<td style=\"background-color: #ffffff;\">GTTGTTTAGTCGAGAGGGTATC<\/td>\n<\/tr>\n<tr>\n<td style=\"text-align: center; background-color: #ffffff;\" rowspan=\"2\">Cat<\/td>\n<td style=\"background-color: #ffffff;\">FW<\/td>\n<td style=\"background-color: #ffffff;\">S39_Cat_F2<\/td>\n<td style=\"background-color: #ffffff;\">TTATCACACCCACAAGAGGA<\/td>\n<td style=\"text-align: center; background-color: #ffffff;\" rowspan=\"2\">1355<\/td>\n<td style=\"text-align: center; background-color: #ffffff;\" rowspan=\"2\">NC 001700<\/td>\n<\/tr>\n<tr>\n<td style=\"background-color: #ffffff;\">RV<\/td>\n<td style=\"background-color: #ffffff;\">S40_Cat_R2<\/td>\n<td style=\"background-color: #ffffff;\">GTAGTACTTTCGACTGGTTAG<\/td>\n<\/tr>\n<tr>\n<td style=\"background-color: #ffffff;\"><\/td>\n<td style=\"background-color: #ffffff;\"><\/td>\n<td style=\"background-color: #ffffff;\"><\/td>\n<td style=\"background-color: #ffffff;\"><\/td>\n<td style=\"background-color: #ffffff;\"><\/td>\n<td style=\"background-color: #ffffff;\"><\/td>\n<\/tr>\n<tr>\n<td style=\"text-align: center; background-color: #ffffff;\" rowspan=\"2\">\u5185\u90e8\u30b3\u30f3\u30c8\u30ed\u30fc\u30eb<br \/>\n18SrRNA<\/td>\n<td style=\"background-color: #ffffff;\">FW<\/td>\n<td style=\"background-color: #ffffff;\">I01_IC_F<\/td>\n<td style=\"background-color: #ffffff;\">CGGGGAATYAGGGTTCGATTC<\/td>\n<td style=\"text-align: center; background-color: #ffffff;\" rowspan=\"2\">70<\/td>\n<td style=\"text-align: center; background-color: #ffffff;\" rowspan=\"2\"><\/td>\n<\/tr>\n<tr>\n<td style=\"background-color: #ffffff;\">RV<\/td>\n<td style=\"background-color: #ffffff;\">I02_IC_R<\/td>\n<td style=\"background-color: #ffffff;\">GCCTGCTGCCTTCCTTKGATG<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<p>\uff0a\u5168\u3066\u306e\u30d7\u30e9\u30a4\u30de\u30fc\u306e\u5e0c\u91c8\u306fTE\u3067\u884c\u3046\u3002<\/p>\n<p>(4)\u8a66\u85ac<\/p>\n<ul>\n<li>Ex Taq Hot Start Version\uff08 TaKaRa Cat#RR006A \uff09<br \/>\n(\u300010\u00d7Ex Taq Buffer \u30012.5mM dNTP Mix \u4ed8\u5c5e\u3000)<\/li>\n<li>TE Buffer\uff08 10mM Tris-HCl(pH8.0) \u30011mM EDTA (pH8.0) \uff09<\/li>\n<li>2% \u30a2\u30ac\u30ed\u30fc\u30b9\u30b2\u30eb<\/li>\n<li>50\u00d7TAE Buffer\uff082M Tris, 2M\u6c37\u9162\u9178, 0.5M EDTA (pH8.0)\uff09<br \/>\n\u4f7f\u7528\u6642\u3001\u7cbe\u88fd\u6c34\u306750\u500d\u5e0c\u91c8\u3059\u308b\u3002\uff081\u00d7TAE\uff09<\/li>\n<li>\u7cbe\u88fd\u6c34<\/li>\n<li>10\u00d7Loading Buffer<\/li>\n<li>DNA\u30b5\u30a4\u30ba\u30de\u30fc\u30ab\u30fc\uff08 100bp DNA Ladder \uff09<\/li>\n<li>10mg\/ml\u30a8\u30c1\u30b8\u30a6\u30e0\u30d6\u30ed\u30de\u30a4\u30c9\u6eb6\u6db2<br \/>\n10mg\/mL\u30a8\u30c1\u30b8\u30a6\u30e0\u30d6\u30ed\u30de\u30a4\u30c910\u03bcL\u3092TAE Buffer 100mL\u306b\u52a0\u3048\u308b\u3002<\/li>\n<\/ul>\n<p><strong>2.\u65b9\u6cd5<\/strong><br \/>\n(1)PCR<\/p>\n<p>1) PCR mixture\u3092\u8abf\u6574\u3057\u3001PCR\u752896\u30a6\u30a7\u30eb\u30d7\u30ec\u30fc\u30c8( \u307e\u305f\u306f0.2ml\u30c1\u30e5\u30fc\u30d6 )\u306b46\u03bcl\u305a\u3064\u5206\u6ce8\u3059\u308b\u3002<\/p>\n<table style=\"margin-left: 40px;\">\n<tbody>\n<tr>\n<td id=\"t1\">\u7cbe\u88fd\u6c34<\/td>\n<td id=\"t1\" style=\"text-align: right; width: 200px;\">32.5 \u03bcl<\/td>\n<\/tr>\n<tr>\n<td id=\"t1\">10\u00d7 Ex Taq buffer<\/td>\n<td id=\"t1\" style=\"text-align: right; width: 200px;\">5 \u03bcl<\/td>\n<\/tr>\n<tr>\n<td id=\"t1\">2.5 mM dNP<\/td>\n<td id=\"t1\" style=\"text-align: right; width: 200px;\">4 \u03bcl<\/td>\n<\/tr>\n<tr>\n<td id=\"t1\">\u52d5\u7269\u7a2e\u7279\u7570\u7684\u30d7\u30e9\u30a4\u30de\u30fcFW (10pmol\/ul)<\/td>\n<td id=\"t1\" style=\"text-align: right; width: 200px;\">1 \u03bcl<\/td>\n<\/tr>\n<tr>\n<td id=\"t1\">\u52d5\u7269\u7a2e\u7279\u7570\u7684\u30d7\u30e9\u30a4\u30de\u30fcRW (10pmol\/ul)<\/td>\n<td id=\"t1\" style=\"text-align: right; width: 200px;\">1 \u03bcl<\/td>\n<\/tr>\n<tr>\n<td id=\"t1\">\u5185\u90e8\u30b3\u30f3\u30c8\u30ed\u30fc\u30eb\u30d7\u30e9\u30a4\u30de\u30fcFW (2pmol\/ul)<\/td>\n<td id=\"t1\" style=\"text-align: right; width: 200px;\">1 \u03bcl<\/td>\n<\/tr>\n<tr>\n<td id=\"t1\">\u5185\u90e8\u30b3\u30f3\u30c8\u30ed\u30fc\u30eb\u30d7\u30e9\u30a4\u30de\u30fcRW (2pmol\/ul)<\/td>\n<td id=\"t1\" style=\"text-align: right; width: 200px;\">1 \u03bcl<\/td>\n<\/tr>\n<tr>\n<td id=\"t1\">Ex Taq HS<\/td>\n<td id=\"t1\" style=\"text-align: right; width: 200px;\">0.25 \u03bcl<\/td>\n<\/tr>\n<tr>\n<td id=\"t1\" colspan=\"2\">\n<hr \/>\n<\/td>\n<\/tr>\n<tr>\n<td id=\"t1\">Total<\/td>\n<td id=\"t1\" style=\"text-align: right;\">46 \u03bcl<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<p>2) \u9670\u6027\u5bfe\u7167\u306f\u30011)\u306ePCR\u752896\u30a6\u30a7\u30eb\u30d7\u30ec\u30fc\u30c8( \u307e\u305f\u306f0.2ml\u30c1\u30e5\u30fc\u30d6 )\u306b\u7cbe\u88fd\u6c344ul\u3092\u52a0\u3048\u308b\u3002<br \/>\n3) \u691c\u67fb\u7d30\u80de\u682a\u304b\u3089\u62bd\u51fa\u3057\u305f\u30b2\u30ce\u30e0DNA\u53ca\u3073\u967d\u6027\u5bfe\u7167\u306f\u3001\u7cbe\u88fd\u6c34\u306725ng\/\u03bcl\u306b\u306a\u308b\u3088\u3046\u306b\u5e0c\u91c8\u3057\u30011)\u306e96\u30a6\u30a7\u30eb\u30d7\u30ec\u30fc\u30c8( \u307e\u305f\u306f0.2ml\u30c1\u30e5\u30fc\u30d6 )\u306b4\u03bcl\u52a0\u3048\u308b\u3002<br \/>\n4) \u652a\u62cc\u51e6\u7406\u3092\u884c\u3044\u3001\u30b9\u30d4\u30f3\u30c0\u30a6\u30f3\u3092\u884c\u3046\u3002<br \/>\n5) PCR program<br \/>\nStage1 : 94\u2103 5\u5206\uff08 1\u30b5\u30a4\u30af\u30eb \uff09<br \/>\nStage2 : 94\u2103 45\u79d2 \u2192 60\u2103 30\u79d2 \u2192 72\u2103 1.5\u5206 ( 30\u30b5\u30a4\u30af\u30eb )<br \/>\nStage3 : 72\u2103 10\u5206\uff08 1\u30b5\u30a4\u30af\u30eb \uff09<br \/>\nStage4 : 4\u2103\uff08 \u4fdd\u5b58 \uff09<\/p>\n<p>(2)\u96fb\u6c17\u6cf3\u52d5<br \/>\n<img decoding=\"async\" src=\"\/ja\/wp-content\/uploads\/images\/pcr_1.png\" alt=\"\u96fb\u6c17\u6cf3\u52d5\" \/><\/p>\n<p><strong>3.\u5224\u5b9a<\/strong><br \/>\n\u9670\u6027\u5bfe\u7167\u306b\u306f\u30d0\u30f3\u30c9\u304c\u306a\u3044\u3053\u3068\u3001\u9670\u6027\u5bfe\u7167\u4ee5\u5916\u306b\u306f\u5185\u90e8\u30b3\u30f3\u30c8\u30ed\u30fc\u30eb\uff0870bp\uff09\u306e\u30d0\u30f3\u30c9\u304c\u51fa\u3066\u3044\u308b\u3053\u3068\u3001\u967d\u6027\u5bfe\u7167\u306b\u30d7\u30e9\u30a4\u30de\u30fc\u4e00\u89a7\u306b\u8a18\u8f09\u306e\u30b5\u30a4\u30ba\u306e\u30d0\u30f3\u30c9\u304c\u51fa\u3066\u3044\u308b\u3053\u3068\u3092\u78ba\u8a8d\u5f8c\u3001\u52d5\u7269\u7a2e\u306e\u5224\u5b9a\u3092\u3059\u308b\u3002<br \/>\n\u3059\u3079\u3066\u306e\u967d\u6027\u5bfe\u7167\u306b\u30d0\u30f3\u30c9\u304c\u51fa\u3066\u3044\u306a\u3044\u5834\u5408\u3001\u5185\u90e8\u30b3\u30f3\u30c8\u30ed\u30fc\u30eb\u306e\u30d0\u30f3\u30c9\u304c\u51fa\u3066\u3044\u306a\u3044\u5834\u5408\u306f\u3001\u518d\u5ea6PCR\u3092\u884c\u3046\u3002<\/p>\n<table style=\"margin-left: 30px;\">\n<tbody>\n<tr>\n<td id=\"t1\">\u967d\u6027\uff08\uff0b\uff09<\/td>\n<td id=\"t1\">\u967d\u6027\u5bfe\u7167\u3068\u540c\u3058\u30b5\u30a4\u30ba\u306e\u30d0\u30f3\u30c9\u304c\u691c\u51fa\u3055\u308c\u305f\u3068\u304d\u306f\u967d\u6027\uff08\uff0b\uff09\u3068\u3057\u3001\u967d\u6027\u5bfe\u7167\u3068\u540c\u3058\u52d5\u7269\u7a2e\u3068\u5224\u5b9a\u3059\u308b\u3002<\/td>\n<\/tr>\n<tr>\n<td id=\"t1\">\u9670\u6027\uff08\uff0d\uff09<\/td>\n<td id=\"t1\">\u967d\u6027\u5bfe\u7167\u3068\u540c\u3058\u30b5\u30a4\u30ba\u306e\u30d0\u30f3\u30c9\u304c\u691c\u51fa\u3055\u308c\u306a\u3044\u3068\u304d\u306f\u9670\u6027\uff08\uff0d\uff09\u3068\u3059\u308b\u3002<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<p>\uff0a\u76ee\u7684\u3068\u3059\u308b\u52d5\u7269\u7a2e\u4ee5\u5916\u306e\u52d5\u7269\u7a2e\u3067\u30d0\u30f3\u30c9\u304c\u691c\u51fa\u3055\u308c\u305f\u5834\u5408\u306f\u3001\u691c\u67fb\u7d30\u80de\u682a\u306b\u4ed6\u7a2e\u7d30\u80de\u304c\u6df7\u5165\u3057\u3066\u3044\u308b\u53ef\u80fd\u6027\u3068\u3001\u7d30\u80de\u306e\u7531\u6765\u304c\u76ee\u7684\u3068\u3059\u308b\u52d5\u7269\u7a2e\u4ee5\u5916\u3067\u3042\u308b\u3053\u3068\u304c\u7591\u308f\u308c\u308b\u3002<\/p>\n","protected":false},"excerpt":{"rendered":"<p>\u7d30\u80de\u682a\u306e\u7531\u6765\u52d5\u7269\u7a2e\u306e\u540c\u5b9a\u3068\u5224\u5225\u53ef\u80fd\u306a\u52d5\u7269\u7a2e\uff08\u6ce81\uff09\u306e\u6df7\u5165\u306a\u3069\u306e\u30af\u30ed\u30b9\u30b3\u30f3\u30bf\u30df\u30cd\u30fc\u30b7\u30e7\u30f3\u306e\u6709\u7121\u3092\u78ba\u8a8d\u3059\u308b\u305f\u3081\u3001\u52d5\u7269\u7a2e\u3092\u540c\u5b9a\u3059\u308bPCR\u691c\u67fb\u3092\u3057\u3066\u3044\u307e\u3059\u3002\u73fe\u5728\u3001\u7d30\u80de\u57f9\u990a\u306b\u5e83\u7bc4\u306b\u7528\u3044\u3089\u308c\u308b\u52d5\u7269\u7a2e13\u7a2e\u306b\u3064\u3044\u3066\u52d5\u7269\u7a2e\u306e\u5224\u5b9a\u304c\u53ef\u80fd\u3067\u3059\u3002 \u6ce81\uff09\u5224\u5225\u53ef\u80fd\u306a\u52d5\u7269\u7a2e \u30d2\u30c8\u3001\u30de\u30a6\u30b9\u3001\u30e9\u30c3\u30c8\u3001\u30cb\u30ef\u30c8\u30ea\u3001\u30b7\u30ea\u30a2\u30f3\u30cf\u30e0\u30b9\u30bf\u30fc\u3001\u30a2\u30d5\u30ea\u30ab\u30df\u30c9\u30ea\u30b6\u30eb\u3001\u30ab\u30cb\u30af\u30a4\u30b6\u30eb\u3001\u30a6\u30b5\u30ae\u3001\u30c1\u30e3\u30a4\u30cb\u30fc\u30ba\u30cf\u30e0\u30b9\u30bf\u30fc\u3001\u30a4\u30cc\u3001\u30d6\u30bf\u3001\u30a6\u30b7\u3001\u30cd\u30b3 \u5185\u90e8\u30b3\u30f3 [&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-10110","page","type-page","status-publish","hentry"],"_links":{"self":[{"href":"http:\/\/cell.brc.riken.jp\/ja\/wp-json\/wp\/v2\/pages\/10110","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=10110"}],"version-history":[{"count":3,"href":"http:\/\/cell.brc.riken.jp\/ja\/wp-json\/wp\/v2\/pages\/10110\/revisions"}],"predecessor-version":[{"id":10865,"href":"http:\/\/cell.brc.riken.jp\/ja\/wp-json\/wp\/v2\/pages\/10110\/revisions\/10865"}],"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=10110"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}