{"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\/news\/lyot-2sddnazbsjhzdzb\/","title":{"rendered":"\u5229\u7528OT-2\u4e0a\u7684\u00aeDNA\u5236\u5907\u8bd5\u5242\u76d2\u81ea\u52a8\u5236\u5907\u5c0f\u57fa\u56e0\u7ec4\u7684DNA\u6587\u5e93"},"content":{"rendered":"\n

\u4f5c\u8005
\u9a6c\u4fee\u00b7\u963f\u5361\u7eb3\uff0c\u91d1\u7eb3\u91cc\u00b7\u6c83\u68ee\u535a\u58eb\uff0c\u83ab\u62c9\u7ea6\u00b7\u963f\u5fb7\u4f0a\u6bd4\u535a\u58eb\u3002<\/p>\n\n\n\n

\u4ecb\u7ecd
\u4e0b\u4e00\u4ee3\u6d4b\u5e8f\uff08NGS\uff09\u7684\u5de5\u4f5c\u6d41\u7a0b\u5305\u62ec\u4f7f\u7528\u5e02\u552e\u8f6f\u4ef6\u5236\u5907DNA\u6587\u5e93\u4ee5\u53ca\u7ecf\u6d4e\u6709\u6548\u7684DNA\u5236\u5907\u8bd5\u5242\u76d2\u3002\u901a\u8fc7\u4f7f\u7528\u81ea\u52a8\u5316\u7684OT-2\u534f\u8bae\u6765\u4f18\u5316\u8fd9\u4e2a\u5de5\u4f5c\u6d41\u5f88\u6709\u5e2e\u52a9\u4ee5\u63d0\u9ad8DNA\u6587\u5e93\u7684\u5236\u5907\u6548\u7387\u3002
\u5728\u8fd9\u91cc\uff0c\u6211\u4eec\u63cf\u8ff0\u4e86\u4f0a\u5229\u00aeDNA\u7684\u6548\u7387\u3002\u51c6\u5907\u5de5\u5177\u5305(Illumina\uff0cCatNo\u300220018705)\u5728OT-2\uff0c\u673a\u5668\u4eba\u6db2\u4f53\u5904\u7406\u5e73\u53f0\u4e0a\uff0c\u81ea\u52a8\u6267\u884c\u6807\u8bb0\u5de5\u4f5c\u6d41\u7a0b\u3002\u4f7f\u7528OT-2\u7684\u9ad8\u6027\u80fd\u80fd\u529b\u81ea\u52a8\u5316\u6b64\u8fc7\u7a0b\u63d0\u4f9b\u4e86\u4e00\u4e2a\u5feb\u901f\u548c\u5065\u58ee\u7684\u5de5\u4f5c\u6d41\u7a0b\uff0c\u751f\u6210\u7528\u4e8e\u57fa\u56e0\u7ec4\u6d4b\u5e8f\u7684\u7edf\u4e00DNA\u51c6\u5907\u6587\u5e93\u3002<\/p>\n\n\n\n

\u7167\u660e\u00aeDNA\u51c6\u5907\u8bd5\u5242\u76d2\u548c\u5916\u4e2d\u5fc3OT-2\u5e73\u53f0\u6982\u8ff0
\u4f0a\u5229\u00ae\u4e13\u5229NGS\u6807\u8bb0\u5de5\u4f5c\u6d41\u7a0b\u5b9e\u73b0\u4e86\u5229\u7528\u73e0\u5b50\u8fdb\u884c\u7684\u73e0\u4e0a\u6807\u8bb0\u94fe\u63a5\u7684\u8f6c\u5ea7\u4f53\u3002\u73e0\u94fe\u7684\u5316\u5b66\u6027\u8d28\u8f6c\u5ea7\u4f53\u5bf9DNA\u7247\u6bb5\u5316\u66f4\u6709\u6548\u548cDNA\u672b\u7aef\u4fee\u590d\u4e0e\u4e4b\u524d\u7684\u6eb6\u6db2\u5185\u6807\u8bb0\u5de5\u4f5c\u6d41\u7a0b\u76f8\u6bd4\u3002\u8be5DNA\u7247\u6bb5\u4f7f\u7528\u53cc\u7d22\u5f15\u9002\u914d\u5668\u8fdb\u884c\u6807\u8bb0\uff0c\u8fd9\u662f\u4e00\u79cd\u72ec\u7279\u7684\u6761\u5f62\u7801\u5206\u914d\u6bcf\u4e2aDNA\u7247\u6bb5\u3002DNA Prep\u6837\u672c\uff08n=8\uff09\u5206\u522b\u4e3a\u91cf\u5316\uff0c\u5f52\u4e00\u5316\u5230\u4e00\u4e2a4nM\u7684\u5e93\uff0c\u6c47\u96c6\u5230\u4e00\u4e2a\u5355\u4e2a\u591a\u8def\u590d\uff088\u500d\u621616\u500d\uff09\u6837\u54c1\uff0c\u7136\u540e\u5728Illumina\u00aeMiSeq\u4e0a\u76842×75\u6d41\u7ec6\u80de\u4e0a\u8fdb\u884c\u57fa\u56e0\u7ec4\u6d4b\u5e8f\u3002\u6211\u4eec\u7684\u6570\u636e\u663e\u793a\u4e86\u6240\u6709\u6570\u636e\u7684\u4f4e\u53ef\u53d8\u6027\uff0cPNA\u6269\u589e\u524d\u540e\u6d4b\u5b9a\u7684DNA\u5236\u5907\u6837\u672c\uff1b\u6761\u5f62\u7801\u5e73\u8861\uff0c\u4ee5\u53ca\u7edf\u4e00\u548c\u9ad8\u8986\u76d6\u7387\u7684\u6d4b\u5e8f\u6bd4\u5bf9\u3002\u53c2\u8003\u57fa\u56e0\u7ec4\u7684\u7814\u7a76\u8868\u660e\uff0c\u4f7f\u7528OT-2\u534f\u8bae\u7684\u81ea\u52a8\u6807\u8bb0\u4e3a\u57fa\u56e0\u7ec4\u6d4b\u5e8f\u63d0\u4f9b\u4e86\u4e00\u4e2a\u9ad8\u8d28\u91cf\u7684DNA\u51c6\u5907\u6587\u5e93\u3002<\/p>\n\n\n\n

OT-2\u5de5\u4f5c\u6d41\u534f\u8bae\u7684\u539f\u7406\u56fe
DNA\u548cIDT\u00ae\u5bf9\u00aeDNA\/RNA UD\u6307\u6570\u96c6A\u7684DNA\u548cIDT\u00ae\u300220027213)\u5728\u5b75\u80b2\u671f\u95f4\u4e00\u4e2a\u88ab\u79f0\u4e3a\u6807\u8bb0\u7684\u8fc7\u7a0b\uff0c\u5728OT-2\u4e0a\uff0c\u5982\u56fe\uff08\u56fe1\uff09\u6240\u793a\u3002\u8fdb\u884c\u4e86\u6807\u8bb0\u6e05\u6d17\u6b65\u9aa4\uff0c\u4ee5\u53bb\u9664\u6b8b\u7559\u7684DNA\u548c\u9002\u914d\u5668\u3002\u4e0b\u4e00\u4e2a\u7684\u7528OT-2\u65b9\u6cd5\u8fdb\u884cDNA\u6269\u589e\u5177\u6709\u53ef\u7f16\u7a0b\u76d6\u548c\u5757\u6e29\u5ea6\u7684\u7532\u677f\u4e0a\u70ed\u5faa\u73af\u5668\u3002<\/p>\n\n\n\n

\"Illumina
\u56fe1. Illumina DNA\u51c6\u5907\u5de5\u4f5c\u6d41\u7a0b\u3002<\/strong>\u865a\u7ebf\u6846\u8868\u793a\u5728OT-2\u4e0a\u81ea\u52a8\u5316\u7684Illumina DNA\u51c6\u5907\u5de5\u4f5c\u6d41\u7a0b\u7684\u6b65\u9aa4<\/em><\/figcaption><\/figure>\n\n\n\n

\u6309\u7167OT-2\u65b9\u6848\uff0c\u8fdb\u884c\u540e\u6269\u589e
\u4f7f\u7528AMPure(\u8d1d\u514b\u66fc\u5e93\u5c14\u7279\uff0c\u732b\u3002\u4e0da63881)\u6839\u636e\u5236\u9020\u5546\u8fdb\u884c\u68c0\u6d4b\u63a8\u8350DNA Prep\u6837\u672c\u88ab\u5b9a\u91cf\uff0c\u5f52\u4e00\u5316\u81f34nM(\u4f7f\u7528NEBNext\u00ae\u6587\u5e93\u5b9a\u91cf\u8bd5\u5242\u76d2\u5bf9\u4f0a\u5229\u00ae\u6587\u5e93\u5b9a\u91cf\u8fdb\u884cqPCR\u6d4b\u5b9a\u8bd5\u5242\u76d2\uff0c\u4ee58\u500d\u621616\u500d\u7684\u5f62\u5f0f\u6c47\u96c6\u6210\u5355\u4e2a\u6837\u672c\uff0c\u7136\u540e\u5728\u4f0a\u5229\u00aeMiSeq\u4e0a\u76842×75\u6d41\u6c60\u4e0a\u8fd0\u884c\u3002\u6839\u636e\u4f0a\u5229\u00aeDNA\/RNA\u72ec\u7279\u53cc\uff08UD\uff09\u7d22\u5f15\u9002\u914d\u5668\u6761\u5f62\u7801\u5e8f\u5217\uff0c\u63d0\u53d6\u6d4b\u5e8f\u6570\u636e\u5e76\u8fdb\u884c\u89e3\u590d\u7528\u3002\u4e0e\u53c2\u8003\u57fa\u56e0\u7ec4\u548c\u8986\u76d6\u56fe\u8c31\u5bf9\u9f50\u7684\u5e8f\u5217\u662f\u901a\u8fc7\u771f\u771f\u54c12021.2.2<\/p>\n\n\n\n

\u5de5\u4f5c\u6d41\u5e03\u5c40
OT-2\u5e73\u53f0\u7684\u5e03\u5c40\u5305\u62ec\u6a21\u5757\uff0c\u5b9e\u9a8c\u5ba4\u8f6f\u4ef6\uff0c\u548c\u4f0a\u5229\u7c73\u7eb3\u00aeDNA\u51c6\u5907\u8bd5\u5242\uff0c\u5982\u8be6\u89c1\uff08\u56fe2\uff09\u3002\u867d\u7136\u8be5\u5de5\u4f5c\u6d41\u5305\u62ec\uff1a\u5728OT-2\u4e0a\u8fdb\u884c\u81ea\u52a8\u79fb\u6db2\uff0c\u4eba\u5de5\u5e72\u9884\u662f\u9700\u8981\u5728\u7532\u677f\u4e0a\u7684\u4e4b\u95f4\u79fb\u52a8\u6837\u54c1\u677f\u70ed\u5faa\u73af\u7b14\u548c\u78c1\u5757\uff0c\u5e76\u5728\u534f\u8bae\u671f\u95f4\u91cd\u7f6e\u79fb\u6db2\u7ba1\u5c16\u7aef\u67b6\u3002<\/p>\n\n\n\n

DNA\u5236\u5907\u6837\u672c\u7684PCR\u6269\u589e
\u91c7\u7528\u4f0a\u5229\u00aeDNA\u5236\u5907\u8bd5\u5242\u76d2\u6784\u5efaOT-2\u4e0a\u7684Lambda DNA\u6587\u5e93\uff08n=8,100 ng\uff09\u3002\u5206\u522b\u5b9a\u91cf\u524d\u540ePCR\u6d53\u5ea6\uff08ng\/uL\uff095\u4e2a\u5468\u671f\u7684PCR\u6269\u589e\uff0c\u548cDNA\u6d53\u5ea6\u4f7f\u7528Quant-iT\u2122dsDNA\u9ad8\u7075\u654f\u5ea6\u68c0\u6d4b\u8bd5\u5242\u76d2(\u70ed\u8d39\u96ea\u79d1\u5b66\u516c\u53f8\uff0cCat\u3002No.Q33120)\u5728\u4e00\u4e2aQubit\u00ae\u4e0a\u786e\u5b9a\u53d8\u5f02\u7cfb\u6570\uff0c\u5373\u6807\u51c6\u5dee\u9664\u4ee5\u6574\u4e2a\u6837\u672c\u7684\u5e73\u5747\u503c\u3002Lambda DNA\u6837\u672c\u7684PCR\u524d\u5e73\u5747\u6d53\u5ea6\u4e3a2.92\uff0c\u53d8\u5f02\u7cfb\u6570\uff08CV\uff09\u4e3a10.4%\uff0c5\u4e2a\u5468\u671f\u540e\u7684PCR\u540e\u6d53\u5ea6\u5e73\u5747\u4e3a28.23\uff0cCV\u4e3a10.7%\uff0c\u89c1\uff08\u88681\uff09\u3002\u8fd9\u4e9b\u7ed3\u679c\u5f88\u7b80\u5355\uff0c\u4f18\u5316\u540e\u7684OT-2\u534f\u8bae\u53ef\u4ee5\u4ea7\u751f\u4e00\u4e2a\u9ad8\u8d28\u91cf\u548c\u53ef\u9760\u7684NGS DNA\u51c6\u5907\u5e93\u3002<\/p>\n\n\n\n

DNA\u5236\u5907\u6587\u5e93\u7684\u5c0f\u57fa\u56e0\u7ec4\u6d4b\u5e8f
\u5bf996\u4e2a\u6837\u672c\u7684\u00aeDNA\u5236\u5907\u8bd5\u5242\u76d2(M)\u8fdb\u884c\u6807\u8bb0\u3002\u7528No. 20018705)\u5236\u5907\u4e86\u5927\u80a0\u6746\u83cc\u975e\u7532\u57fa\u5316\u57fa\u56e0\u7ec4DNA (Zymogen\uff0cCat\u3002. \u4e0d. \u6837\u54c1\uff08n=16,100ng\uff09<\/p>\n\n\n\n

\"\u7532\u677f\u5e03\u5c40\u914d\u5907\u6a21\u5757\u3001\u5b9e\u9a8c\u5ba4\u5668\u76bf\u548cIllumina
\u56fe2.OT-2\u7532\u677f\u5e03\u5c40\u914d\u5907\u6a21\u5757\u3001\u5b9e\u9a8c\u5ba4\u5668\u76bf\u548cIllumina DNA\u51c6\u5907\u8bd5\u5242\u3002<\/figcaption><\/figure>\n\n\n\n

\u8fd9\u4e2a\u5de5\u4f5c\u6d41\u5728OT-2\u4e0a\u90fd\u6709\u81ea\u52a8\u7684\u79fb\u6db2\u529f\u80fd\uff0c\u9700\u8981\u624b\u52a8\u64cd\u4f5c\u5728\u7532\u677f\u4e0a\u79fb\u52a8\u6837\u54c1\u677f(1)\u548c\u78c1\u6027\u78c1\u5757(2)\uff0c\u4ee5\u53ca\u590d\u4f4d\u5728\u8fd0\u884c\u671f\u95f4\u7684\u5c16\u7aef\u673a\u67b6\u3002\u7532\u677f\u5e03\u5c40\u5305\u62ec1\u4e2aNEST 12\u4e95\u6c34\u5e93\uff0c1\u4e2a96\u4e95\u94dd\u5757\uff0c1\u500d\u57c3\u5f6d\u591a\u592b\u70ed\u5faa\u73af\u5668\u4e0a\u7684200ul PCR\u677f\uff0c\u548c\u4e00\u4e2a1xNEST0.1 mL PCR\u677f\u4e0a\u6e29\u5ea6\u6a21\u5757(3)\u3002\u6a21\u5757\u5305\u62ec\u4e00\u4e2a\u78c1\u6027\u6a21\u5757\u3001\u6e29\u5ea6\u6a21\u5757\u3001\u70ed\u5faa\u73af\u5668\u6a21\u5757\uff0c\u4ee5\u53caP20\u548cP300\u591a\u901a\u9053\u79fb\u6db2\u7ba1\u3002<\/p>\n\n\n\n

\"\u5b9e\u9a8c\u7ed3\u679c\u6570\u636e\"
\u88681.\u4e24\u8005\u4e4b\u95f4\u7684\u4f4e\u53d8\u5f02\u6027\u5e93\u6784\u5efa\u5728Opentrons OT-2.<\/strong>\u8be5\u8868\u663e\u793a\u4e86\u6bcf\u4e2a\u6837\u54c1\u5728Illumina\u6807\u7b7e\u6e05\u6d17\u540e\u7684\u6536\u7387\u7ecf\u8fc75\u4e2a\u5468\u671f\u7684PCR\u6269\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\u7ec4\u7684\u6d4b\u5e8f\u6279\u6b21\u89c4\u683c\u663e\u793a\u51fa\u4f4e\u53d8\u5f02\u3002<\/strong>\u7528Illumina DNA\u5236\u5907\u5de5\u4f5c\u6d41\u7a0b\u5728OT-2\u4e0a\u5236\u5907\u76843\u4e2a\u4e0d\u540c\u6279\u6b21\u7684\u6bd4\u8f83(1)\u3002\u6765\u81ea\u4e0d\u540c\u6837\u54c1\u7c7b\u578b\u7684\u4ea7\u91cf\u662f\u4e00\u81f4\u7684(2)\uff0c\u5355\u72ec\u7684\u6837\u672c\u6761\u5f62\u7801\u4e5f\u663e\u793a\u4e86\u7edf\u4e00\u7684\u8868\u793a\u65b9\u5f0f(3)\u3002\u5e73\u5747\u6837\u672c\u57fa\u56e0\u7ec4\u8986\u76d6\u7387\u8fd1\u4f3c\u4e8e\u4eceIllumina\u8986\u76d6\u7387\u8ba1\u7b97\u5668\u8ba1\u7b97\u51fa\u7684\u9884\u671f\u8986\u76d6\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\"
\u56fe3.\u5728\u5c0f\u57fa\u56e0\u7ec4\u4e2d\u5177\u6709\u9ad8\u5ea6\u4e00\u81f4\u7684\u6d4b\u5e8f\u8986\u76d6\u7387\u3002<\/strong>\u6765\u81ea\u57fa\u56e0\u7ec4\u7684\u8986\u76d6\u56fe\u663e\u793a~7800x\u7684Lambda\u57fa\u56e0\u7ec4\u8986\u76d648kb<\/em><\/figcaption><\/figure>\n\n\n\n

\u4e24\u6279\uff0c\u7b2c\u4e8c\u6279\uff0cE\u3002. \u5927\u80a0\u6746\u83ccDNA\u6837\u672c
\uff08n=8,100 ng\uff09\u5728OT-2\u4e0a\u76842\u4e2a\u5355\u72ec\u7684\u5217\u4e0a\u8fdb\u884c\u5904\u7406\uff0c\u4ee5\u89e3\u91ca\u5217\u5185\u53ef\u80fd\u51fa\u73b0\u7684\u53ef\u53d8\u6027\u3002
E. \u5927\u80a0\u6746\u83ccDNA\uff08n=16,100 ng\uff09\u548cLambda DNA (n=8,100ng)\u88ab\u6269\u589e\u3001\u5f52\u4e00\u5316\uff0c\u5e76\u6c47\u96c6\u5230\u4e00\u4e2a\u5355\u4e00\u7684\u591a\u8def\u590d\u7528\uff088\u500d\u621616\u500d\uff09\u6837\u672c\u4e2d\u8fdb\u884c\u6d4b\u5e8f\u3002
\u5bf9\u4e09\u4e2aDNA\u5236\u5907\u6279\u6b21\u7684\u6d4b\u5e8f\u7ed3\u679c\u4e3a\u6bd4\u8f83\u786e\u5b9a\u6837\u54c1\u4ea7\u91cf\uff0c\u6761\u5f62\u7801
. \u5e73\u8861\u548c\u8986\u76d6\u7387\uff0c\u5982\uff08\u88682\uff09\u6240\u793a\uff0c\u5bf9DNA\u5236\u5907\u6587\u5e93\u89c2\u5bdf\u5230\u7edf\u4e00\u7684\u4ea7\u91cf\u3002\u8ba1\u7b97\u4e86E\u7684\u6837\u672c\u4ea7\u91cf\u30028\u53f7\u8bd5\u9a8c\u5ba4\u7684\u5927\u80a0\u6746\u83cc\u542b\u91cf\u5206\u522b\u4e3a9.75\u548c3.79
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