{"id":3037,"date":"2024-06-28T11:17:38","date_gmt":"2024-06-28T03:17:38","guid":{"rendered":"https:\/\/opentrons.com.cn\/?post_type=knowledge2&p=3037"},"modified":"2024-09-02T18:30:49","modified_gmt":"2024-09-02T10:30:49","slug":"lyot-2sddnazbsjhzdzb","status":"publish","type":"post","link":"https:\/\/opentrons.com.cn\/zh-TW\/news\/lyot-2sddnazbsjhzdzb\/","title":{"rendered":"\u5229\u7528OT-2\u4e0a\u7684\u00aeDNA\u88fd\u5099\u8a66\u5291\u76d2\u81ea\u52d5\u88fd\u5099\u5c0f\u57fa\u56e0\u7d44\u7684DNA\u6587\u5eab"},"content":{"rendered":"\n

\u4f5c\u8005\u99ac\u4fee\u2027\u963f\u5361\u7d0d\uff0c\u91d1\u7d0d\u88e1\u2027\u6c83\u68ee\u535a\u58eb\uff0c\u83ab\u62c9\u7d04\u2027\u963f\u5fb7\u4f0a\u6bd4\u535a\u58eb\u3002<\/p>\n\n\n\n

\u4ecb\u7d39\u6b21\u4e16\u4ee3\u5b9a\u5e8f\uff08NGS\uff09\u7684\u5de5\u4f5c\u6d41\u7a0b\u5305\u62ec\u4f7f\u7528\u5e02\u552e\u8edf\u9ad4\u88fd\u5099DNA\u6587\u5eab\u4ee5\u53ca\u7d93\u6fdf\u6709\u6548\u7684DNA\u88fd\u5099\u8a66\u5291\u76d2\u3002\u900f\u904e\u4f7f\u7528\u81ea\u52d5\u5316\u7684OT-2\u5354\u8b70\u4f86\u512a\u5316\u9019\u500b\u5de5\u4f5c\u6d41\u7a0b\u5f88\u6709\u5e6b\u52a9\u4ee5\u63d0\u9ad8DNA\u6587\u5eab\u7684\u88fd\u5099\u6548\u7387\u3002\u5728\u9019\u88e1\uff0c\u6211\u5011\u63cf\u8ff0\u4e86\u4f0a\u5229\u00aeDNA\u7684\u6548\u7387\u3002\u6e96\u5099\u5de5\u5177\u5305(Illumina\uff0cCatNo\u300220018705)\u5728OT-2\uff0c\u6a5f\u5668\u4eba\u6db2\u9ad4\u8655\u7406\u5e73\u53f0\u4e0a\uff0c\u81ea\u52d5\u57f7\u884c\u6a19\u8a18\u5de5\u4f5c\u6d41\u7a0b\u3002\u4f7f\u7528OT-2\u7684\u9ad8\u6027\u80fd\u80fd\u529b\u81ea\u52d5\u5316\u6b64\u904e\u7a0b\u63d0\u4f9b\u4e86\u4e00\u500b\u5feb\u901f\u4e14\u5065\u58ef\u7684\u5de5\u4f5c\u6d41\u7a0b\uff0c\u7522\u751f\u7528\u65bc\u57fa\u56e0\u7d44\u5b9a\u5e8f\u7684\u7d71\u4e00DNA\u6e96\u5099\u6587\u5eab\u3002<\/p>\n\n\n\n

\u7167\u660e\u00aeDNA\u6e96\u5099\u8a66\u5291\u76d2\u548c\u5916\u4e2d\u5fc3OT-2\u5e73\u53f0\u6982\u8ff0\u4f0a\u5229\u00ae\u5c08\u5229NGS\u6a19\u8a18\u5de5\u4f5c\u6d41\u7a0b\u5be6\u73fe\u4e86\u5229\u7528\u73e0\u5b50\u9032\u884c\u7684\u73e0\u4e0a\u6a19\u8a18\u93c8\u63a5\u7684\u8f49\u5ea7\u9ad4\u3002\u73e0\u93c8\u7684\u5316\u5b78\u6027\u8cea\u8f49\u5ea7\u9ad4\u5c0dDNA\u7247\u6bb5\u5316\u66f4\u6709\u6548\u548cDNA\u672b\u7aef\u4fee\u5fa9\u8207\u5148\u524d\u6eb6\u6db2\u5167\u6a19\u8a18\u7684\u5de5\u4f5c\u6d41\u7a0b\u76f8\u6bd4\u3002\u6b64DNA\u7247\u6bb5\u4f7f\u7528\u96d9\u7d22\u5f15\u9069\u914d\u5668\u9032\u884c\u6a19\u8a18\uff0c\u9019\u662f\u4e00\u7a2e\u7368\u7279\u7684\u689d\u78bc\u5206\u914d\u6bcf\u500bDNA\u7247\u6bb5\u3002 DNA Prep\u6a23\u672c\uff08n=8\uff09\u5206\u5225\u70ba\u91cf\u5316\uff0c\u6b78\u4e00\u5316\u5230\u4e00\u500b4nM\u7684\u5eab\uff0c\u532f\u96c6\u5230\u4e00\u500b\u55ae\u4e00\u591a\u8def\u5fa9\uff088\u500d\u621616\u500d\uff09\u6a23\u54c1\uff0c\u7136\u5f8c\u5728Illumina\u00aeMiSeq\u4e0a\u76842x75\u6d41\u7d30\u80de\u4e0a\u9032\u884c\u57fa\u56e0\u7d44\u6e2c\u5e8f\u3002\u6211\u5011\u7684\u6578\u64da\u986f\u793a\u4e86\u6240\u6709\u6578\u64da\u7684\u4f4e\u8b8a\u7570\u6027\uff0cPNA\u64f4\u589e\u524d\u5f8c\u6e2c\u5b9a\u7684DNA\u88fd\u5099\u6a23\u672c\uff1b\u689d\u78bc\u5e73\u8861\uff0c\u4ee5\u53ca\u7d71\u4e00\u548c\u9ad8\u8986\u84cb\u7387\u7684\u5b9a\u5e8f\u6bd4\u5c0d\u3002\u53c3\u8003\u57fa\u56e0\u7d44\u7684\u7814\u7a76\u8868\u660e\uff0c\u4f7f\u7528OT-2\u5354\u8b70\u7684\u81ea\u52d5\u6a19\u8a18\u70ba\u57fa\u56e0\u7d44\u6e2c\u5e8f\u63d0\u4f9b\u4e86\u4e00\u500b\u9ad8\u54c1\u8cea\u7684DNA\u6e96\u5099\u6587\u5eab\u3002<\/p>\n\n\n\n

OT-2\u5de5\u4f5c\u6d41\u7a0b\u5354\u8b70\u7684\u539f\u7406\u5716DNA\u548cIDT\u00ae\u5c0d\u00aeDNA\/RNA UD\u6307\u6578\u96c6A\u7684DNA\u548cIDT\u00ae\u3002 20027213)\u5728\u5b75\u80b2\u671f\u9593\u4e00\u500b\u88ab\u7a31\u70ba\u6a19\u8a18\u7684\u904e\u7a0b\uff0c\u5728OT-2\u4e0a\uff0c\u5982\u5716\uff08\u57161\uff09\u6240\u793a\u3002\u9032\u884c\u4e86\u6a19\u8a18\u6e05\u6d17\u6b65\u9a5f\uff0c\u4ee5\u53bb\u9664\u6b98\u7559\u7684DNA\u548c\u9069\u914d\u5668\u3002\u4e0b\u4e00\u500b\u7684\u7528OT-2\u65b9\u6cd5\u9032\u884cDNA\u64f4\u589e\u5177\u6709\u53ef\u7de8\u7a0b\u84cb\u548c\u584a\u6eab\u5ea6\u7684\u7532\u677f\u4e0a\u71b1\u5faa\u74b0\u5668\u3002<\/p>\n\n\n\n

\"Illumina
\u57161. Illumina DNA\u6e96\u5099\u5de5\u4f5c\u6d41\u7a0b\u3002 <\/strong>\u865b\u7dda\u6846\u8868\u793a\u5728OT-2\u4e0a\u81ea\u52d5\u5316\u7684Illumina DNA\u6e96\u5099\u5de5\u4f5c\u6d41\u7a0b\u7684\u6b65\u9a5f<\/em><\/figcaption><\/figure>\n\n\n\n

\u4f9d\u7167OT-2\u65b9\u6848\uff0c\u9032\u884c\u5f8c\u64f4\u589e\u4f7f\u7528AMPure(\u8c9d\u514b\u66fc\u5eab\u723e\u7279\uff0c\u8c93\u3002\u4e0da63881)\u6839\u64da\u88fd\u9020\u5546\u9032\u884c\u6aa2\u6e2c\u63a8\u85a6DNA Prep\u6a23\u672c\u88ab\u5b9a\u91cf\uff0c\u6b78\u4e00\u5316\u81f34nM(\u4f7f\u7528NEBNext\u00ae\u6587\u5eab\u5b9a\u91cf\u8a66\u5291\u76d2\u5c0d\u4f0a\u5229\u00ae\u6587\u5eab\u5b9a\u91cf\u9032\u884cqPCR\u6e2c\u5b9a\u8a66\u5291\u76d2\uff0c\u4ee58\u500d\u621616\u500d\u7684\u5f62\u5f0f\u532f\u96c6\u55ae\u4e00\u6a23\u672c\uff0c\u7136\u5f8c\u5728\u4f0a\u5229\u00aeMiSeq\u4e0a\u76842x75\u6d41\u6c60\u4e0a\u904b\u884c\u3002<\/p>\n\n\n\n

\u5de5\u4f5c\u6d41\u7a0b\u4f48\u5c40OT-2\u5e73\u53f0\u7684\u4f48\u7f6e\u5305\u62ec\u6a21\u7d44\uff0c\u5be6\u9a57\u5ba4\u8edf\u9ad4\uff0c\u548c\u4f0a\u5229\u7c73\u7d0d\u00aeDNA\u6e96\u5099\u8a66\u5291\uff0c\u5982\u8a73\u898b\uff08\u57162\uff09\u3002\u96d6\u7136\u8a72\u5de5\u4f5c\u6d41\u7a0b\u5305\u62ec\uff1a\u5728OT-2\u4e0a\u9032\u884c\u81ea\u52d5\u79fb\u6db2\uff0c\u4eba\u5de5\u5e79\u9810\u662f\u9700\u8981\u5728\u7532\u677f\u4e0a\u7684\u4e4b\u9593\u79fb\u52d5\u6a23\u54c1\u677f\u71b1\u5faa\u74b0\u7b46\u548c\u78c1\u584a\uff0c\u4e26\u5728\u5354\u8b70\u671f\u9593\u91cd\u7f6e\u79fb\u6db2\u7ba1\u5c16\u7aef\u67b6\u3002<\/p>\n\n\n\n

DNA\u88fd\u5099\u6a23\u672c\u7684PCR\u64f4\u589e\u63a1\u7528\u4f0a\u5229\u00aeDNA\u88fd\u5099\u8a66\u5291\u76d2\u5efa\u69cbOT-2\u4e0a\u7684Lambda DNA\u6587\u5eab\uff08n=8,100 ng\uff09\u3002\u5206\u5225\u5b9a\u91cf\u524d\u5f8cPCR\u6fc3\u5ea6\uff08ng\/uL\uff095\u500b\u9031\u671f\u7684PCR\u64f4\u589e\uff0c\u548cDNA\u6fc3\u5ea6\u4f7f\u7528Quant-iT\u2122dsDNA\u9ad8\u9748\u654f\u5ea6\u6aa2\u6e2c\u8a66\u5291\u76d2(\u71b1\u8cbb\u96ea\u79d1\u5b78\u516c\u53f8\uff0cCat\u3002No.Q33120)\u5728\u4e00\u500bQubit\u00ae\u4e0a\u78ba\u5b9a\u8b8a\u7570\u4fc2\u6578\uff0c\u5373\u6a19\u6e96\u5dee\u9664\u4ee5\u6574\u500b\u6a23\u672c\u7684\u5e73\u5747\u503c\u3002 Lambda DNA\u6a23\u672c\u7684PCR\u524d\u5e73\u5747\u6fc3\u5ea6\u70ba2.92\uff0c\u8b8a\u7570\u4fc2\u6578\uff08CV\uff09\u70ba10.4%\uff0c5\u500b\u9031\u671f\u5f8c\u7684PCR\u5f8c\u6fc3\u5ea6\u5e73\u5747\u70ba28.23\uff0cCV\u70ba10.7%\uff0c\u898b\uff08\u88681\uff09\u3002\u9019\u4e9b\u7d50\u679c\u5f88\u7c21\u55ae\uff0c\u512a\u5316\u5f8c\u7684OT-2\u5354\u8b70\u53ef\u4ee5\u7522\u751f\u4e00\u500b\u9ad8\u54c1\u8cea\u548c\u53ef\u9760\u7684NGS DNA\u6e96\u5099\u5eab\u3002<\/p>\n\n\n\n

DNA\u88fd\u5099\u6587\u5eab\u7684\u5c0f\u57fa\u56e0\u7d44\u5b9a\u5e8f\u5c0d96\u500b\u6a23\u672c\u7684\u00aeDNA\u88fd\u5099\u8a66\u5291\u76d2(M)\u9032\u884c\u6a19\u8a18\u3002\u7528No. 20018705)\u88fd\u5099\u4e86\u5927\u8178\u687f\u83cc\u975e\u7532\u57fa\u5316\u57fa\u56e0\u7d44DNA (Zymogen\uff0cCat.. \u4e0d. \u6a23\u672c\uff08n=16,100ng\uff09<\/p>\n\n\n\n

\"\u7532\u677f\u5e03\u5c40\u914d\u5907\u6a21\u5757\u3001\u5b9e\u9a8c\u5ba4\u5668\u76bf\u548cIllumina
\u57162.OT-2\u7532\u677f\u4f48\u7f6e\u914d\u5099\u6a21\u7d44\u3001\u5be6\u9a57\u5ba4\u5668\u76bf\u548cIllumina DNA\u6e96\u5099\u8a66\u5291\u3002<\/figcaption><\/figure>\n\n\n\n

\u9019\u500b\u5de5\u4f5c\u6d41\u7a0b\u5728OT-2\u4e0a\u90fd\u6709\u81ea\u52d5\u7684\u79fb\u6db2\u529f\u80fd\uff0c\u9700\u8981\u624b\u52d5\u64cd\u4f5c\u5728\u7532\u677f\u4e0a\u79fb\u52d5\u6a23\u54c1\u677f(1)\u548c\u78c1\u6027\u78c1\u584a(2)\uff0c\u4ee5\u53ca\u5fa9\u4f4d\u5728\u904b\u4f5c\u671f\u9593\u7684\u5c16\u7aef\u6a5f\u67b6\u3002\u7532\u677f\u4f48\u5c40\u5305\u62ec1\u500bNEST 12\u4e95\u6c34\u5eab\uff0c1\u500b96\u4e95\u92c1\u584a\uff0c1\u500d\u57c3\u5f6d\u591a\u592b\u71b1\u5faa\u74b0\u5668\u4e0a\u7684200ul PCR\u677f\uff0c\u548c\u4e00\u500b1xNEST0.1 mL PCR\u677f\u4e0a\u6eab\u5ea6\u6a21\u7d44(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\u591a\u901a\u9053\u79fb\u6db2\u7ba1\u3002<\/p>\n\n\n\n

\"\u5b9e\u9a8c\u7ed3\u679c\u6570\u636e\"
\u88681.\u5169\u8005\u4e4b\u9593\u7684\u4f4e\u8b8a\u7570\u6027\u5eab\u69cb\u5efa\u5728Opentrons OT-2.<\/strong>\u8a72\u8868\u986f\u793a\u4e86\u6bcf\u500b\u6a23\u54c1\u5728Illumina\u6a19\u7c64\u6e05\u6d17\u5f8c\u7684\u6536\u7387\u7d93\u904e5\u500b\u9031\u671f\u7684PCR\u64f4\u589e\u548cAMPure\u6e05\u9664<\/em><\/figcaption><\/figure>\n\n\n\n
\"\u5c0f\u57fa\u56e0\u7ec4\u7684\u6d4b\u5e8f\u6279\u6b21\u89c4\u683c\u663e\u793a\u51fa\u4f4e\u53d8\u5f02\"
\u88682.\u5c0f\u57fa\u56e0\u7d44\u7684\u5b9a\u5e8f\u6279\u6b21\u898f\u683c\u986f\u793a\u4f4e\u8b8a\u7570\u3002 <\/strong>\u4ee5Illumina DNA\u88fd\u5099\u5de5\u4f5c\u6d41\u7a0b\u5728OT-2\u4e0a\u88fd\u5099\u76843\u500b\u4e0d\u540c\u6279\u6b21\u7684\u6bd4\u8f03(1)\u3002\u4f86\u81ea\u4e0d\u540c\u6a23\u54c1\u985e\u578b\u7684\u7522\u91cf\u662f\u4e00\u81f4\u7684(2)\uff0c\u55ae\u7368\u7684\u6a23\u672c\u689d\u78bc\u4e5f\u986f\u793a\u4e86\u7d71\u4e00\u7684\u8868\u793a\u65b9\u5f0f(3)\u3002\u5e73\u5747\u6a23\u672c\u57fa\u56e0\u7d44\u8986\u84cb\u7387\u8fd1\u4f3c\u65bc\u5f9eIllumina\u8986\u84cb\u7387\u8a08\u7b97\u5668\u8a08\u7b97\u51fa\u7684\u9810\u671f\u8986\u84cb\u7387\u3002 <\/em><\/figcaption><\/figure>\n\n\n\n
\"\u5728\u5c0f\u57fa\u56e0\u7ec4\u4e2d\u5177\u6709\u9ad8\u5ea6\u4e00\u81f4\u7684\u6d4b\u5e8f\u8986\u76d6\u7387\"
\u57163.\u5728\u5c0f\u57fa\u56e0\u7d44\u4e2d\u5177\u6709\u9ad8\u5ea6\u4e00\u81f4\u7684\u5b9a\u5e8f\u8986\u84cb\u7387\u3002 <\/strong>\u4f86\u81ea\u57fa\u56e0\u7d44\u7684\u8986\u84cb\u5716\u986f\u793a~7800x\u7684Lambda\u57fa\u56e0\u7d44\u8986\u84cb48kb<\/em><\/figcaption><\/figure>\n\n\n\n

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