{"id":257,"date":"2020-12-02T12:09:46","date_gmt":"2020-12-02T03:09:46","guid":{"rendered":"https:\/\/www.promega.co.jp\/resources\/faq\/?p=257"},"modified":"2021-04-07T12:07:13","modified_gmt":"2021-04-07T03:07:13","slug":"q83","status":"publish","type":"post","link":"https:\/\/www.promega.co.jp\/resources\/faq\/amplification-quantification\/pcr-and-rt\/hignfidelity\/pfu-dna-poly\/q83","title":{"rendered":"\u306a\u305cTaq DNA Polymerase\u3067\u306f\u306a\u304f\u3001Pfu DNA Polymerase\u304c\u4f7f\u308f\u308c\u308b\u306e\u3067\u3059\u304b? Pfu DNA Polymerase\u306b\u3088\u308b\u53cd\u5fdc\u306f\u3069\u306e\u3088\u3046\u306b\u8abf\u88fd\u3059\u308c\u3070\u3044\u3044\u306e\u3067\u3059\u304b?"},"content":{"rendered":"\n

Pfu DNA Polymerase(\u30ab\u30bf\u30ed\u30b0\u756a\u53f7 M7741)\u306f\u3001\u9ad8\u5fe0\u5b9f\u5ea6\u304c\u5fc5\u8981\u3068\u306a\u308bPCR\u3067\u4f7f\u7528\u3059\u308b\u3001Pyrococcus furiosus Vc1 (DSM3638) (1)\u304b\u3089\u5358\u96e2\u3055\u308c\u305f\u8010\u71b1\u6027\u9175\u7d20\u3067\u3059\u3002\u3053\u306e\u9175\u7d20\u306f5\u2019\u21923\u2019\u306eDNA\u30dd\u30ea\u30e1\u30e9\u30fc\u30bc\u6d3b\u6027\u306b\u52a0\u3048\u3066\u30013\u2019\u21925\u2019\u306e\u30a8\u30ad\u30bd\u30cc\u30af\u30ec\u30a2\u30fc\u30bc\u6d3b\u6027(\u30d7\u30eb\u30fc\u30d5\u30ea\u30fc\u30c7\u30a3\u30f3\u30b0\u6d3b\u6027)\u3082\u793a\u3057\u307e\u3059\u3002Pfu DNA Polymerase\u306f\u3053\u308c\u307e\u3067\u306b\u660e\u3089\u304b\u306b\u306a\u3063\u3066\u3044\u308b\u8010\u71b1\u6027\u9175\u7d20\u306e\u4e2d\u3067\u6700\u3082\u4f4e\u3044\u30a8\u30e9\u30fc\u7387\u3092\u793a\u3057\u307e\u3059(2-4)\u3002\u305d\u306e\u305f\u3081\u3001Pfu DNA Polymerase\u306f\u9ad8\u3044\u5fe0\u5b9f\u5ea6\u3092\u5fc5\u8981\u3068\u3059\u308bDNA\u5408\u6210\u306b\u5f79\u7acb\u3061\u307e\u3059(2-5)\u3002\u30dd\u30ea\u30e1\u30e9\u30fc\u30bc\u306e\u6b63\u78ba\u3055\u3068\u306f\u3001\u305d\u306e\u30dd\u30ea\u30e1\u30e9\u30fc\u30bc\u304c\u8aa4\u3063\u305f\u30cc\u30af\u30ec\u30aa\u30c1\u30c9\u3092\u53d6\u308a\u8fbc\u3080\u524d\u307e\u3067\u306b\u53d6\u308a\u8fbc\u307e\u308c\u305f\u30cc\u30af\u30ec\u30aa\u30c1\u30c9\u306e\u6570\u3067\u3059\u3002\u30a8\u30e9\u30fc\u7387\u306f\u6b63\u78ba\u3055\u306e\u9006\u306e\u5024\u3067\u3001\u8907\u88fd\u3055\u308c\u305f\u30d9\u30fc\u30b9\u30da\u30a2\u3042\u305f\u308a\u306e\u5909\u7570\u6570\u3067\u8868\u3055\u308c\u307e\u3059\u3002Pfu DNA Polymerase\u306e\u6b63\u78ba\u3055\u306f7.7 x 105(4)\uff5e1 x 106(6)\u3068\u5831\u544a\u3055\u308c\u3066\u3044\u307e\u3059\u3002\u540c\u69d8\u306ePCR\u3092\u5143\u306b\u3057\u305f\u8a66\u9a13\u3067Pfu DNA Polymerase\u306f\u3001Vent\u00ae DNA Polymerase (New England Biolabs, Inc.)\u306e\u7d042\u500d\u3001Taq DNA Polymerase (3,4)\u306e\u7d046\u500d\u306e\u6b63\u78ba\u3055\u3092\u793a\u3057\u307e\u3059\u3002\u9ad8\u3044\u5fe0\u5b9f\u5ea6\u3092\u5f97\u308b\u305f\u3081\u306b\u306f\u3001\u5897\u5e45\u53cd\u5fdc\u306b\u4f7f\u3046Tris\u7cfb\u306e10\u00d7\u30d0\u30c3\u30d5\u30a1\u30fc\u306epH\u306f8.6\u4ee5\u4e0a(25\u2103)\u3067\u306a\u3051\u308c\u3070\u306a\u308a\u307e\u305b\u3093\u3002pH8.0\u3067\u306fPfu DNA Polymerase\u306e\u5fe0\u5b9f\u5ea6\u306fTaq DNA Polymarase\u306e\u5fe0\u5b9f\u5ea6\u3088\u308a\u3082\u52a3\u308a\u307e\u3059\u3002\u30d7\u30ed\u30e1\u30ac\u306ePfu DNA Polymerase\u306b\u6dfb\u4ed8\u3055\u308c\u308b\u53cd\u5fdc\u30d0\u30c3\u30d5\u30a1\u30fc\u306epH\u306f\u9ad8\u5fe0\u5b9f\u5ea6\u3092\u5b9f\u73fe\u3059\u308bpH8.8\u3067\u3059\u3002\u3053\u306e\u53cd\u5fdc\u30d0\u30c3\u30d5\u30a1\u30fc\u306b\u306f200\u03bcM\u306e\u5404dNTP\u3092\u53cd\u5fdc\u306b\u4f7f\u3046\u3068\u304d\u306b\u6700\u9069\u306a\u9175\u7d20\u6d3b\u6027\u3092\u793a\u3059\u3088\u3046\u306b2mM MgSO4\u3082\u542b\u307e\u308c\u3066\u3044\u307e\u3059\u3002\u53cd\u5fdc\u6db2\u306b\u52a0\u3048\u308b\u92f3\u578bDNA\u306e\u91cf\u3092\u5897\u3084\u3059\u3053\u3068\u3067\u5fe0\u5b9f\u5ea6\u3092\u4e0a\u3052\u308b\u3053\u3068\u304c\u3067\u304d\u307e\u3059\u3002\u307e\u305f\u3001\u30b5\u30a4\u30af\u30eb\u6570\u3092\u6e1b\u3089\u3059\u3053\u3068\u3084\u5909\u6027\u306e\u6e29\u5ea6\u4e0b\u306b\u304a\u304f\u6642\u9593\u3092\u5c11\u306a\u304f\u3059\u308b\u3053\u3068\u3067\u3082\u53ef\u80fd\u3067\u3059\u3002\u3053\u306e\u3088\u3046\u306bPCR\u7523\u7269\u306e\u7c21\u5358\u306a\u691c\u51fa\u306e\u3088\u3046\u306a\u4e00\u822c\u7684\u306aPCR\u306b\u306fTaq DNA Polymerase\u3084Tfl DNA Polymerase\u304c\u9069\u3057\u3066\u3044\u307e\u3059\u3002\u4e00\u65b9\u3001\u907a\u4f1d\u5b50\u306e\u30af\u30ed\u30fc\u30cb\u30f3\u30b0\u3084\u767a\u73fe\u3001\u5909\u7570\u306e\u89e3\u6790\u306e\u3088\u3046\u306a\u9ad8\u5fe0\u5b9f\u5ea6\u304c\u5fc5\u8981\u3068\u306a\u308bPCR\u306b\u306f\u3001Pfu DNA Polymerase\u3092\u63a8\u5968\u3057\u307e\u3059\u3002

Pfu<\/em> DNA Polymerase\u3092\u5897\u5e45\u53cd\u5fdc\u6db2\u3078\u6700\u5f8c\u306b\u52a0\u3048\u308b\u3053\u3068(\u7279\u306bdNTPs\u3092\u52a0\u3048\u305f\u5f8c\u306b)\u304c\u91cd\u8981\u3067\u3059\u3002\u3053\u308c\u306fPfu<\/em> DNA Polymerase\u306e\u30d7\u30eb\u30fc\u30d5\u30ea\u30fc\u30c7\u30a3\u30f3\u30b0\u6d3b\u6027\u306b\u3088\u308a\u30d7\u30e9\u30a4\u30de\u30fc\u3092\u5206\u89e3\u3057\u3001\u975e\u7279\u7570\u7684\u306a\u5897\u5e45\u7523\u7269\u304c\u73fe\u308c\u305f\u308a\u3001\u53ce\u91cf\u306e\u4f4e\u4e0b\u3092\u8d77\u3053\u3059\u53ef\u80fd\u6027\u304c\u3042\u308b\u304b\u3089\u3067\u3059\u3002\u30d7\u30e9\u30a4\u30de\u30fc\u306e\u5206\u89e3\u3092\u9632\u3050\u305f\u3081\u306bGC\u542b\u91cf\u306e\u9ad8\u3044\u9577\u3081\u306e\u30d7\u30e9\u30a4\u30de\u30fc\u3092\u4f7f\u3063\u305f\u5834\u5408\u30015\u2019\u5074\u306e\u521d\u3081\u306e15\u5869\u57fa\u306f\u5206\u89e3\u304b\u3089\u307b\u307c\u5b8c\u5168\u306b\u4fdd\u8b77\u3055\u308c\u307e\u3059\u3002\u3057\u305f\u304c\u3063\u3066\u9069\u5207\u306a\u30d7\u30e9\u30a4\u30de\u30fc\u306e\u9577\u3055\u306f25\uff5e30\u5869\u57fa\u3067\u3059\u3002\u307e\u305f\u3001\u30d7\u30e9\u30a4\u30de\u30fc\u306e3\u2019\u672b\u7aef\u3078\u306e\u30db\u30b9\u30db\u30ed\u30c1\u30aa\u30a8\u30fc\u30c8(phosphorothioate)\u306e\u4ed8\u52a0\u306b\u3088\u308a\u4fdd\u8b77\u3059\u308b\u3053\u3068\u3082\u3067\u304d\u307e\u3059(7)\u3002\u5897\u5e45\u53cd\u5fdc\u6db2\u306f\u6c37\u4e0a\u3067\u6e96\u5099\u3057\u300195\u2103\u306b\u3042\u3089\u304b\u3058\u3081\u71b1\u3057\u3066\u304a\u3044\u305f\u30b5\u30fc\u30de\u30eb\u30b5\u30a4\u30af\u30e9\u30fc\u306b\u30bb\u30c3\u30c8\u3059\u308b\u3053\u3068\u3092\u63a8\u5968\u3057\u307e\u3059\u3002\u4f38\u9577\u53cd\u5fdc\u306f1kb\u3042\u305f\u308a2\u5206\u9593\u3067\u884c\u3063\u3066\u304f\u3060\u3055\u3044\u3002\uff08eNotes-FAQ0004)<\/p>\n\n\n\n

\u53c2\u7167\u6587\u732e:<\/strong><\/p>\n\n\n\n

  1. Fiala, G. and Stetter, K.O. (1986) Pyrococcus furiosus sp. nov. represents a novel genus of marine heterotrophic archaebacteria growing optimally at 100\u2103. Arch. Microbiol. 145, 56.<\/li>
  2. Flaman, J.M. et al. (1994) A rapid PCR fidelity assay. Nucl. Acids Res. 22, 3259.<\/li>
  3. Andre, P. et al. (1997) Fidelity and mutational spectrum of Pfu DNA polymerase on a human mitochondrial DNA sequence. Genome Res. 7, 843.<\/li>
  4. Cline, J., Braman, J.C. and Hogrefe, H.H. (1996) PCR fidelity of Pfu DNA polymerase and other thermostable DNA polymerases. Nucl. Acids Res. 24, 3546.<\/li>
  5. Lundberg, K.S. et al. (1991) High-fidelity amplification using a thermostable DNA polymerase isolated from Pyrococcus furiosus. Gene 108, 1.<\/li>
  6. Slater, M. et al. (1998) Pfu DNA Polymerase: A High Fidelity Enzyme for Nucleic Acid Amplification. Promega Notes 68, 7.<\/li>
  7. Skerra, A. (1992) Phosphorothioate primers improve the amplification of DNA sequences by DNA polymerases with proofreading activity. Nucl. Acids Res. 20, 3551.<\/li><\/ol>\n","protected":false},"excerpt":{"rendered":"

    Pfu DNA Polymerase(\u30ab\u30bf\u30ed\u30b0\u756a\u53f7 M7741)\u306f\u3001\u9ad8\u5fe0\u5b9f\u5ea6\u304c\u5fc5\u8981\u3068\u306a\u308bPCR\u3067\u4f7f\u7528\u3059\u308b\u3001Pyrococcus furiosus Vc1 (DSM3638) (1)\u304b\u3089\u5358\u96e2\u3055\u308c\u305f\u8010\u71b1\u6027\u9175\u7d20\u3067\u3059\u3002\u3053\u306e\u9175\u7d20\u306f … <\/p>\n","protected":false},"author":15,"featured_media":0,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":[],"categories":[91],"tags":[],"_links":{"self":[{"href":"https:\/\/www.promega.co.jp\/resources\/faq\/wp-json\/wp\/v2\/posts\/257"}],"collection":[{"href":"https:\/\/www.promega.co.jp\/resources\/faq\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/www.promega.co.jp\/resources\/faq\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/www.promega.co.jp\/resources\/faq\/wp-json\/wp\/v2\/users\/15"}],"replies":[{"embeddable":true,"href":"https:\/\/www.promega.co.jp\/resources\/faq\/wp-json\/wp\/v2\/comments?post=257"}],"version-history":[{"count":3,"href":"https:\/\/www.promega.co.jp\/resources\/faq\/wp-json\/wp\/v2\/posts\/257\/revisions"}],"predecessor-version":[{"id":2703,"href":"https:\/\/www.promega.co.jp\/resources\/faq\/wp-json\/wp\/v2\/posts\/257\/revisions\/2703"}],"wp:attachment":[{"href":"https:\/\/www.promega.co.jp\/resources\/faq\/wp-json\/wp\/v2\/media?parent=257"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/www.promega.co.jp\/resources\/faq\/wp-json\/wp\/v2\/categories?post=257"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/www.promega.co.jp\/resources\/faq\/wp-json\/wp\/v2\/tags?post=257"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}