{"id":3014,"date":"2024-06-25T23:33:08","date_gmt":"2024-06-25T15:33:08","guid":{"rendered":"https:\/\/opentrons.com.cn\/?post_type=knowledge2&#038;p=3014"},"modified":"2024-09-02T18:57:10","modified_gmt":"2024-09-02T10:57:10","slug":"sydnazbsjhzdzb","status":"publish","type":"post","link":"https:\/\/opentrons.com.cn\/zh-TW\/news\/sydnazbsjhzdzb\/","title":{"rendered":"\u4f7f\u7528DNA\u88fd\u5099\u8a66\u5291\u76d2\u81ea\u52d5\u88fd\u5099\u5c0f\u57fa\u56e0\u7d44\u7684\u6587\u5eab"},"content":{"rendered":"<!-- wp:paragraph -->\n<p>\u6458\u8981\u4e09\u500b\u5716\u66f8\u9928\u6e96\u5099\u5de5\u5177\u5305-Illumina\u00aeDNA\u6e96\u5099\u5de5\u5177\u5305\uff0cKAPA\u00ae\u8d85\u7d1a\u6e96\u5099\u5de5\u5177\u5305\u548cNEBNext\u00aeUltra\u2122II\u8a66\u5291\u76d2-\u90fd\u662f\u900f\u904eOpentrons 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-->\n<p>\u4ecb\u7d39DNA\u6587\u5eab\u7684\u88fd\u5099\u662f\u4e0b\u4e00\u6b65\u5de5\u4f5c\u7684\u95dc\u9375\u6b65\u9a5f\u4ee3\u5b9a\u5e8f\uff08NGS\uff09\u3002\u81ea\u52d5\u5316\u7684DNA\u6587\u5eab\u88fd\u5099\u70ba\u5efa\u69cb\u9ad8\u7522\u91cf\u5b9a\u5e8f\u6587\u5eab\u63d0\u4f9b\u4e86\u4e00\u7a2e\u7c21\u5316\u7684\u65b9\u6cd5\u540c\u6642\u76e1\u91cf\u6e1b\u5c11\u5be6\u969b\u64cd\u4f5c\u6642\u9593\u3002\u5728\u9019\u88e1\uff0c\u6211\u5011\u63cf\u8ff0\u6d77\u767e\u5408DNA\u88fd\u5099\u81ea\u52d5\u5316\u7684\u6548\u7387\u88dd\u5099(\u4f0a\u5229\uff0cCatNo\u300220018705)\uff0cKAPA\u8d85\u7d1a\u8a66\u5291\u76d2(\u7f85\u6c0f\u00ae. , CatNo.7962363001)\u548cNEBNext\u8d85II\u8a66\u5291\u76d2(\u65b0\u82f1\u683c\u862d\u751f\u7269\u5be6\u9a57\u5ba4\u00ae. , CatNo .E7645L)\u5728OT-2\u6a5f\u5668\u4eba\u6db2\u9ad4\u8655\u7406\u5e73\u53f0\u4e0a\u3002<\/p>\n<!-- \/wp:paragraph -->\n\n<!-- wp:paragraph -->\n<p>\u6750\u6599\u8207\u65b9\u6cd5Illumina DNA\u88fd\u5099\u8a66\u5291\u76d2\u3001KAPA\u8d85\u7d1a\u88fd\u5099\u8a66\u5291\u76d2\u3001NEBNext Ultra II\u548c\u8996\u4e2d\u5fc3OT-2\u5e73\u53f0\u6982\u8ff0\u64c1\u6709\u5c08\u5229NGS\u6a19\u8a18\u5de5\u4f5c\u6d41\u7a0b\uff0c\u5229\u7528\u73e0\u806f\u8f49\u5ea7\u9ad4\uff0c\u9019\u4e0d\u540c\u65bcKAPA HyperPrep\u548cNEBNext Ultra II\u5de5\u4f5c\u6d41\u7a0b-DNA\u672b\u7aef\u4fee\u5fa9\u548ca\u5c3e\u7684\u89e3\u6c7a\u6d41\u7a0b\u3002<\/p>\n<!-- \/wp:paragraph -->\n\n<!-- wp:paragraph -->\n<p>\u5c0d\u65bcIllumina DNA\u6e96\u5099\u8a66\u5291\u76d2\uff0c\u4f7f\u7528OT-2\u4e0a\u7684\u96d9\u7d22\u5f15\u9069\u914d\u5668\u5c0dDNA\u9032\u884c\u6a19\u8a18\u548c\u6a19\u8a18\u3002 DNA\u6a23\u672c\uff08n=8,100 ng\uff09\u91cf\u5316\uff0c\u6b78\u4e00\u5316\u5230\u4e00\u500b4 nM\u7684\u5eab\uff0c\u532f\u96c6\u5230\u4e00\u500b\u591a\u8def\u5fa9\u7528\u7684\u6a23\u672c\uff0c\u7136\u5f8c\u5728\u4e00\u500b2x75\u4e0a\u904b\u884c\u5728Illumina MiSeq\u4e0a\u7684\u6d41\u5f0f\u7d30\u80de\u9032\u884c\u57fa\u56e0\u7d44\u6e2c\u5e8f\u3002\u5de5\u4f5c\u6d41\u7a0b\u5305\u62ec\u7522\u751fDNA\u6e96\u5099\u5eabLambda\uff08n=24\uff09\u548c\u4eba\u985e\u57fa\u56e0\u7d44DNA\u6a23\u672c\uff08n=8\uff09\u3002<\/p>\n<!-- \/wp:paragraph -->\n\n<!-- wp:paragraph -->\n<p>\u5c0d\u65bcHyperPrep\u548cNEBNext Ultra II DNA\u88fd\u5099\u8a66\u5291\u76d2\uff0c\u6211\u5011\u5728OT-2\u4e0a\u5efa\u69cb\u4e86\u6297\u9ad4DNA\u7247\u6bb5\uff08n=8,100 ng\uff09\u7684DNA\u6587\u5eab\u3002\u4f7f\u7528\u7684\u689d\u78bc\u4f86\u81eaKAPA\u7368\u7279\u7684\u96d9\u7d22\u5f15\u9069\u914d\u5668\uff0c\u548cNEB\u591a\u91cd\u96d9\u7d22\u5f15II\u70baNEBNext\u7d22\u5f15\u8a2d\u5b9a1\u3002\u6a23\u672c\u88ab\u5b9a\u91cf\u3001\u6b78\u4e00\u5316\uff0c\u4e26\u5f59\u96c6\u5230\u4e00\u500b4 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\/wp:paragraph -->\n\n<!-- wp:paragraph -->\n<p>OT-2\u5de5\u4f5c\u6d41\u7a0b\u5354\u5b9a\u7684\u539f\u7406\u5716<\/p>\n<!-- \/wp:paragraph -->\n\n<!-- wp:paragraph -->\n<p>\u5c0d\u65bcIllumina\u7684DNA\u6e96\u5099\u8a66\u5291\u76d2\uff0cDNA\u548cIDT\u00ae\u70ba\u4e86\u4f0a\u5229DNA\/RNA UD\u6307\u6578\u96c6A\u3002 .\u5728OT-2\u4e0a\u9032\u884c\u4e86\u6a19\u8a18\uff08\u57161\uff09\u3002 A\u6e05\u6d17\u6b65\u9a5f\u662f\u5fc5\u8981\u7684\uff0c\u4ee5\u53bb\u9664\u6b98\u7559\u7684DNA\u548c\u9069\u914d\u5668\u63a5\u4e0b\u4f86\uff0c\u4f7f\u7528\u5177\u6709\u53ef\u7de8\u7a0b\u84cb\u5b50\u548c\u584a\u6eab\u5ea6\u7684OT-2\u7532\u677f\u4e0a\u71b1\u5faa\u74b0\u5668\u9032\u884cPCR\u64f4\u589e\u3002<\/p>\n<!-- \/wp:paragraph -->\n\n<!-- wp:paragraph -->\n<p>\u5c0d\u65bcHyperPrep\u548cNEBNext Ultra II DNA\u88fd\u5099\u8a66\u5291\u76d2\uff0c\u90fd\u4ee5NEB\u7247\u6bb5\u9175\u7d20\u9032\u884c\u7247\u6bb5\u5316\u3002\u4eba\u985e16\u5206\u9418\uff0c\u4eba\u985e20\u5206\u9418\u57fa\u56e0\u7d44DNA\uff0c\u7136\u5f8c\u7528AMPure\u9032\u884c\u6e05\u7406\u00aeXP\u73e0\u5b50(\u8c9d\u514b\u66fc\u5eab\u723e\u7279\uff0cCatNo\u3002.A63881).\u5c0dOT-2\u9032\u884cDNA\u672b\u7aef\u4fee\u5fa9\/A\u5c3e\u8de1\uff0c\u7136\u5f8c\u662fAMPure XP\u73e0\u5b50\u6e05\u7406\u6b65\u9a5f\u3001\u689d\u78bc\u6216\u9069\u914d\u5668\u9023\u63a5\uff08\u57161\uff09\u3002\u4f7f\u7528\u7684\u689d\u78bc\u4f86\u81eaKAPA\u7368\u7279\u7684\u96d9\u7d22\u5f15\u9069\u914d\u5668(\u7f85\u6c0f\uff0cCatNo\u300208861919702). \u5c0d\u65bcHyperPrep\u548cNEB\u591a\u91cd\u96d9\u7d22\u5f15\u5f15\u5b50\u96c61 (NEB\uff0cCatNo\u3002E7335L)\u70baNEBNext Ultra II\u3002 .<\/p>\n<!-- \/wp:paragraph -->\n\n<!-- wp:paragraph -->\n<p>\u4f7f\u7528\u7532\u677f\u4e0a\u7684\u71b1\u5faa\u74b0\u5668\u5728OT-2\u4e0a\u9032\u884cDNA\u64f4\u589e\u3002<\/p>\n<!-- \/wp:paragraph -->\n\n<!-- wp:paragraph -->\n<p>\u5de5\u4f5c\u6d41\u7a0b\u4f48\u5c40Opentrons OT-2\u5e73\u53f0\u7684\u4f48\u7f6e\u5305\u62ec\u6a21\u7d44\u3001\u5be6\u9a57\u5ba4\u8edf\u9ad4\u548cDNA\u6e96\u5099\u8a66\u5291\uff08\u57162\uff09\u3002\u96d6\u7136\u5de5\u4f5c\u6d41\u7a0b\u5305\u62ec\u5728OT-2\u4e0a\u7684\u81ea\u52d5\u79fb\u6db2\uff0c\u4f46\u9700\u8981\u624b\u52d5\u5e79\u9810\uff0c\u5728\u7532\u677f\u4e0a\u7684\u6046\u6eab\u5668\u548c\u78c1\u584a\u4e4b\u9593\u79fb\u52d5\u6a23\u54c1\u677f\uff0c\u4e26\u5728\u6b64\u671f\u9593\u91cd\u7f6e\u79fb\u6db2\u7ba1\u5c16\u7aef\u67b6\u5354\u8b70\u3002<\/p>\n<!-- \/wp:paragraph -->\n\n<!-- wp:image {\"id\":3015,\"sizeSlug\":\"full\",\"linkDestination\":\"none\"} -->\n<figure class=\"wp-block-image size-full\"><img src=\"https:\/\/opentrons.com.cn\/wp-content\/uploads\/2024\/06\/image-76.png\" alt=\"\u5de5\u4f5c\u6d41\u5e03\u5c40\" class=\"wp-image-3015\"><\/figure>\n<!-- \/wp:image -->\n\n<!-- wp:image {\"id\":3016,\"sizeSlug\":\"full\",\"linkDestination\":\"none\",\"align\":\"left\"} -->\n<figure class=\"wp-block-image alignleft size-full\"><img src=\"https:\/\/opentrons.com.cn\/wp-content\/uploads\/2024\/06\/image-77.png\" alt=\"OT-2\u7532\u677f\u5e03\u5c40\" class=\"wp-image-3016\"><\/figure>\n<!-- \/wp:image -->\n\n<!-- wp:paragraph -->\n<p><strong>\u57162\uff1aOT-2\u7532\u677f\u4f48\u5c40<\/strong><\/p>\n<!-- \/wp:paragraph -->\n\n<!-- wp:paragraph -->\n<p><strong>\u6a21\u7d44\uff0c\u5be6\u9a57\u5ba4\u8edf\u9ad4\uff0c\u548cDNA\u6e96\u5099\u8a66\u5291\u3002\u9019\u500b<\/strong><\/p>\n<!-- \/wp:paragraph -->\n\n<!-- wp:paragraph -->\n<p>\u6a19\u6e96\u5316\u7684\u7532\u677f\u4f48\u5c40\u8207HyperPrep\u548c\u5176\u4ed6\u5e73\u53f0\u5171\u4eab<\/p>\n<!-- \/wp:paragraph -->\n\n<!-- wp:paragraph -->\n<p>NEB\u4e0b\u4e00\u500bUltra II\u3002\u6b64\u5de5\u4f5c\u6d41\u7a0b\u5728OT-2\u4e0a\u81ea\u52d5\u79fb\u6db2\uff0c\u9700\u8981\u624b\u52d5\u5e79\u9810\u5728\u7532\u677f\u4e0a\u6046\u6eab\u7b46\u548c\u4e4b\u9593\u79fb\u52d5\u6a23\u54c1\u677f(1)<\/p>\n<!-- \/wp:paragraph -->\n\n<!-- wp:paragraph -->\n<p>\u78c1\u6027\u78c1\u584a(2)\uff0c\u4ee5\u53ca\u5728\u904b\u4f5c\u904e\u7a0b\u4e2d\u91cd\u7f6e\u5c16\u7aef\u6a5f\u67b6\u3002\u7532\u677f\u4f48\u5c40\u5305\u62ec1\u500bNEST 12\u4e95\u5132\u5c64\uff0c1 x 96\u4e95\u92c1\u584a\uff0c1\u500b\u71b1\u5faa\u74b0\u5668\u4e0a\u7684200\u00b5l PCR\u677f\uff0c\u6eab\u5ea6\u6a21\u7d44\u4e0a\u76841\u500bNEST0.1 mL PCR\u677f(3)\u3002\u6a21\u7d44\u5305\u62ec\u4e00\u500b\u78c1\u6027\u6a21\u7d44\u3001\u6eab\u5ea6\u6a21\u7d44\u3001\u71b1\u5faa\u74b0\u5668\u6a21\u7d44\uff0c\u4ee5\u53caP20\u548cP300 8\u901a\u9053\u79fb\u6db2\u7ba1\u3002<\/p>\n<!-- \/wp:paragraph -->\n\n<!-- wp:paragraph -->\n<p>\u7d50\u679cDNA\u6587\u5eab\u64f4\u589e\u4f7f\u7528PicoGreen\u6aa2\u6e2cDNA\u6587\u5eab\u00ae\u5728\u4e00\u500b\u91cf\u5b50\u4f4d\u5143\u00ae4.\u7528\u87a2\u5149\u8a08\u6e2c\u5b9a\u6fc3\u5ea6\u3002 CV\uff08\u8b8a\u7570\u4fc2\u6578\u70ba\u6a19\u6e96\u5dee\u9664\u4ee5\u5e73\u5747\u503c\uff09\u3002\u6240\u6709\u6587\u5eab\u7684\u6700\u7d42\u6d17\u812b\u9ad4\u7a4d\u5747\u70ba30\u00b5L\u3002\u5c0d\u65bc\u4f7f\u7528Illumina\u6e96\u5099\u7684\u5eabDNAPrep\uff08n=8,100ng\uff09\u7684CV\u70ba10.7%\uff08n=24,100 ng\uff09\u7684CV\u70ba16.6%\uff08\u88681A\uff09\u3002\u70ba\u6587\u5eab\u4f7f\u7528HyperPrep\u88fd\u5099\uff0cCV\u70ba\uff08n=8100ng\uff09\u768413.4%\uff0c\uff08n=24100ng\uff09\u7684CV\u70ba17.6<\/p>\n<!-- \/wp:paragraph -->\n\n<!-- wp:paragraph -->\n<p>. 1B)\u4f7f\u7528NEBNext Ultra 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