{"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\/news\/sydnazbsjhzdzb\/","title":{"rendered":"\u4f7f\u7528DNA\u5236\u5907\u8bd5\u5242\u76d2\u81ea\u52a8\u5236\u5907\u5c0f\u57fa\u56e0\u7ec4\u7684\u6587\u5e93"},"content":{"rendered":"\n<p>\u6458\u8981<br>\u4e09\u4e2a\u56fe\u4e66\u9986\u51c6\u5907\u5de5\u5177\u5305-Illumina\u00aeDNA\u51c6\u5907\u5de5\u5177\u5305\uff0cKAPA\u00ae\u8d85\u7ea7\u51c6\u5907\u5de5\u5177\u5305\u548cNEBNext\u00aeUltra\u2122II\u8bd5\u5242\u76d2-\u90fd\u662f\u901a\u8fc7Opentrons OT-2\u81ea\u52a8\u5feb\u901f\u548c\u7a33\u5065\u5730\u5236\u5907\u4e0b\u4e00\u4ee3\u6d4b\u5e8f\u7684\u9ad8\u8d28\u91cf\u6587\u5e93\u3002<br>\u4ecelambda\u548c\u4eba\u7c7b\u57fa\u56e0\u7ec4DNA\u4e2d\u5236\u5907\u4e86100 ng\u8f93\u5165\u7684\u6587\u5e93\u3002<br>\u89c2\u5bdf\u5230\u5e93\u7684\u7c7b\u4f3c\u6027\u80fd\uff0c\u7528OT-2\u4e0a\u7684\u4e09\u4e2a\u5957\u4ef6\u51c6\u5907\u7684\u6837\u672c\u53d8\u5f02\u6027\u548c\u4ea7\u91cf\u3002\u7c7b\u4f3c\u7684\u6392\u5e8f\u6307\u6807\uff0c\u5982\u6761\u5f62\u7801\u5e73\u8861\u548c\u5e8f\u5217\u6bd4\u5bf9\u4e5f\u89c2\u5bdf\u5230\u8986\u76d6\u8303\u56f4\u3002<\/p>\n\n\n\n<p>\u4ecb\u7ecd<br>DNA\u6587\u5e93\u7684\u5236\u5907\u662f\u4e0b\u4e00\u6b65\u5de5\u4f5c\u7684\u5173\u952e\u6b65\u9aa4\u4ee3\u6d4b\u5e8f\uff08NGS\uff09\u3002\u81ea\u52a8\u5316\u7684DNA\u6587\u5e93\u5236\u5907\u4e3a\u6784\u5efa\u9ad8\u4ea7\u6d4b\u5e8f\u6587\u5e93\u63d0\u4f9b\u4e86\u4e00\u79cd\u7b80\u5316\u7684\u65b9\u6cd5\u540c\u65f6\u5c3d\u91cf\u51cf\u5c11\u5b9e\u9645\u64cd\u4f5c\u65f6\u95f4\u3002\u5728\u8fd9\u91cc\uff0c\u6211\u4eec\u63cf\u8ff0\u6d77\u767e\u5408DNA\u5236\u5907\u81ea\u52a8\u5316\u7684\u6548\u7387\u88c5\u5907(\u4f0a\u5229\uff0cCatNo\u300220018705)\uff0cKAPA\u8d85\u7ea7\u8bd5\u5242\u76d2(\u7f57\u6c0f\u00ae. 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ng\uff09\u91cf\u5316\uff0c\u5f52\u4e00\u5316\u5230\u4e00\u4e2a4 nM\u7684\u5e93\uff0c\u6c47\u96c6\u5230\u4e00\u4e2a\u591a\u8def\u590d\u7528\u7684\u6837\u672c\uff0c\u7136\u540e\u5728\u4e00\u4e2a2&#215;75\u4e0a\u8fd0\u884c\u5728Illumina MiSeq\u4e0a\u7684\u6d41\u5f0f\u7ec6\u80de\u8fdb\u884c\u57fa\u56e0\u7ec4\u6d4b\u5e8f\u3002\u5de5\u4f5c\u6d41\u7a0b\u5305\u62ec\u751f\u6210DNA\u51c6\u5907\u5e93Lambda\uff08n=24\uff09\u548c\u4eba\u7c7b\u57fa\u56e0\u7ec4DNA\u6837\u672c\uff08n=8\uff09\u3002<\/p>\n\n\n\n<p>\u5bf9\u4e8eHyperPrep\u548cNEBNext Ultra II DNA\u5236\u5907\u8bd5\u5242\u76d2\uff0c\u6211\u4eec\u5728OT-2\u4e0a\u6784\u5efa\u4e86\u6297\u4f53DNA\u7247\u6bb5\uff08n=8,100 ng\uff09\u7684DNA\u6587\u5e93\u3002\u4f7f\u7528\u7684\u6761\u5f62\u7801\u6765\u81eaKAPA\u72ec\u7279\u7684\u53cc\u7d22\u5f15\u9002\u914d\u5668\uff0c\u548cNEB\u591a\u91cd\u53cc\u7d22\u5f15II\u4e3aNEBNext\u7d22\u5f15\u8bbe\u7f6e1\u3002\u6837\u672c\u88ab\u5b9a\u91cf\u3001\u5f52\u4e00\u5316\uff0c\u5e76\u6c47\u96c6\u5230\u4e00\u4e2a4 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DNA\u5236\u5907\u8bd5\u5242\u76d2\uff0c\u90fd\u7528NEB\u7247\u6bb5\u9176\u8fdb\u884c\u7247\u6bb5\u5316\u3002<br>\u4eba\u7c7b16\u5206\u949f\uff0c\u4eba\u7c7b20\u5206\u949f\u57fa\u56e0\u7ec4DNA\uff0c\u7136\u540e\u7528AMPure\u8fdb\u884c\u6e05\u7406\u00aeXP\u73e0\u5b50(\u8d1d\u514b\u66fc\u5e93\u5c14\u7279\uff0cCatNo\u3002. A63881).\u5bf9OT-2\u8fdb\u884cDNA\u672b\u7aef\u4fee\u590d\/A\u5c3e\u8ff9\uff0c\u7136\u540e\u662fAMPure XP\u73e0\u5b50\u6e05\u7406\u6b65\u9aa4\u3001\u6761\u5f62\u7801\u6216\u9002\u914d\u5668\u8fde\u63a5\uff08\u56fe1\uff09\u3002\u4f7f\u7528\u7684\u6761\u5f62\u7801\u6765\u81eaKAPA\u72ec\u7279\u7684\u53cc\u7d22\u5f15\u9002\u914d\u5668(\u7f57\u6c0f\uff0cCatNo\u300208861919702). \u5bf9\u4e8eHyperPrep\u548cNEB\u591a\u91cd\u53cc\u7d22\u5f15\u5f15\u7269\u96c61 (NEB\uff0cCatNo\u3002E7335L)\u4e3aNEBNext Ultra II\u3002.<\/p>\n\n\n\n<p>\u4f7f\u7528\u7532\u677f\u4e0a\u7684\u70ed\u5faa\u73af\u5668\u5728OT-2\u4e0a\u8fdb\u884cDNA\u6269\u589e\u3002<\/p>\n\n\n\n<p>\u5de5\u4f5c\u6d41\u5e03\u5c40<br>Opentrons OT-2\u5e73\u53f0\u7684\u5e03\u5c40\u5305\u62ec\u6a21\u5757\u3001\u5b9e\u9a8c\u5ba4\u8f6f\u4ef6\u548cDNA\u51c6\u5907\u8bd5\u5242\uff08\u56fe2\uff09\u3002<br>\u867d\u7136\u5de5\u4f5c\u6d41\u7a0b\u5305\u62ec\u5728OT-2\u4e0a\u7684\u81ea\u52a8\u79fb\u6db2\uff0c\u4f46\u9700\u8981\u624b\u52a8\u5e72\u9884\uff0c\u5728\u7532\u677f\u4e0a\u7684\u6052\u6e29\u5668\u548c\u78c1\u5757\u4e4b\u95f4\u79fb\u52a8\u6837\u54c1\u677f\uff0c\u5e76\u5728\u6b64\u671f\u95f4\u91cd\u7f6e\u79fb\u6db2\u7ba1\u5c16\u7aef\u67b6\u534f\u8bae\u3002<\/p>\n\n\n\n<figure class=\"wp-block-image size-full\"><img decoding=\"async\" width=\"758\" height=\"464\" src=\"https:\/\/opentrons.com.cn\/wp-content\/uploads\/2024\/06\/image-76.png\" alt=\"\u5de5\u4f5c\u6d41\u5e03\u5c40\" class=\"wp-image-3015\" srcset=\"https:\/\/opentrons.com.cn\/wp-content\/uploads\/2024\/06\/image-76.png 758w, https:\/\/opentrons.com.cn\/wp-content\/uploads\/2024\/06\/image-76-300x184.png 300w, https:\/\/opentrons.com.cn\/wp-content\/uploads\/2024\/06\/image-76-576x353.png 576w\" sizes=\"(max-width: 758px) 100vw, 758px\" \/><\/figure>\n\n\n<div class=\"wp-block-image\">\n<figure class=\"alignleft size-full\"><img decoding=\"async\" width=\"474\" height=\"350\" src=\"https:\/\/opentrons.com.cn\/wp-content\/uploads\/2024\/06\/image-77.png\" alt=\"OT-2\u7532\u677f\u5e03\u5c40\" class=\"wp-image-3016\" srcset=\"https:\/\/opentrons.com.cn\/wp-content\/uploads\/2024\/06\/image-77.png 474w, https:\/\/opentrons.com.cn\/wp-content\/uploads\/2024\/06\/image-77-300x222.png 300w\" sizes=\"(max-width: 474px) 100vw, 474px\" \/><\/figure><\/div>\n\n\n<p><strong>\u56fe2\uff1aOT-2\u7532\u677f\u5e03\u5c40<\/strong><\/p>\n\n\n\n<p><strong>\u6a21\u5757\uff0c\u5b9e\u9a8c\u5ba4\u8f6f\u4ef6\uff0c\u548cDNA\u51c6\u5907\u8bd5\u5242\u3002\u8fd9\u4e2a<\/strong><\/p>\n\n\n\n<p>\u6807\u51c6\u5316\u7684\u7532\u677f\u5e03\u5c40\u4e0eHyperPrep\u548c\u5176\u4ed6\u5e73\u53f0\u5171\u4eab<\/p>\n\n\n\n<p>NEB\u4e0b\u4e00\u4e2aUltra II\u3002\u8be5\u5de5\u4f5c\u6d41\u7a0b\u5728OT-2\u4e0a\u81ea\u52a8\u79fb\u6db2\uff0c\u9700\u8981\u624b\u52a8\u5e72\u9884\u5728\u7532\u677f\u4e0a\u6052\u6e29\u7b14\u548c\u4e4b\u95f4\u79fb\u52a8\u6837\u54c1\u677f(1)<\/p>\n\n\n\n<p>\u78c1\u6027\u78c1\u5757(2)\uff0c\u4ee5\u53ca\u5728\u8fd0\u884c\u8fc7\u7a0b\u4e2d\u91cd\u7f6e\u5c16\u7aef\u673a\u67b6\u3002\u7532\u677f\u5e03\u5c40\u5305\u62ec1\u4e2aNEST 12\u4e95\u50a8\u5c42\uff0c1 x 96\u4e95\u94dd\u5757\uff0c1\u4e2a\u70ed\u5faa\u73af\u5668\u4e0a\u7684200\u00b5l PCR\u677f\uff0c\u6e29\u5ea6\u6a21\u5757\u4e0a\u76841\u4e2aNEST0.1 mL PCR\u677f(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 8\u901a\u9053\u79fb\u6db2\u7ba1\u3002<\/p>\n\n\n\n<p>\u7ed3\u679c<br>DNA\u6587\u5e93\u6269\u589e<br>\u4f7f\u7528PicoGreen\u68c0\u6d4bDNA\u6587\u5e93\u00ae\u5728\u4e00\u4e2a\u91cf\u5b50\u4f4d\u00ae4.\u7528\u8367\u5149\u8ba1\u6d4b\u5b9a\u6d53\u5ea6\u3002CV\uff08\u53d8\u5f02\u7cfb\u6570\u4e3a\u6807\u51c6\u5dee\u9664\u4ee5\u5e73\u5747\u503c\uff09\u3002\u6240\u6709\u6587\u5e93\u7684\u6700\u7ec8\u6d17\u8131\u4f53\u79ef\u5747\u4e3a30\u00b5L\u3002\u5bf9\u4e8e\u4f7f\u7528Illumina\u51c6\u5907\u7684\u5e93<br>DNAPrep\uff08n=8,100ng\uff09\u7684CV\u4e3a10.7%\uff08n=24,100 ng\uff09\u7684CV\u4e3a16.6%\uff08\u88681A\uff09\u3002\u4e3a\u6587\u5e93\u4f7f\u7528HyperPrep\u5236\u5907\uff0cCV\u4e3a\uff08n=8100ng\uff09\u768413.4%\uff0c\uff08n=24100ng\uff09\u7684CV\u4e3a17.6<\/p>\n\n\n\n<p>. 1B)\u4f7f\u7528NEBNext Ultra 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