{"id":3063,"date":"2024-06-28T14:04:03","date_gmt":"2024-06-28T06:04:03","guid":{"rendered":"https:\/\/opentrons.com.cn\/?post_type=knowledge2&#038;p=3063"},"modified":"2024-09-04T18:07:35","modified_gmt":"2024-09-04T10:07:35","slug":"xlzbqzdwswhrldnambdzb","status":"publish","type":"knowledge2","link":"https:\/\/zh-tw.opentrons.com.cn\/knowledge2\/xlzbqzdwswhrldnambdzb\/","title":{"rendered":"\u5e8f\u5217\u6e96\u5099\u5168\u81ea\u52d5\u5fae\u751f\u7269\u548c\u4eba\u985eDNA\u6a21\u677f\u7684\u6587\u5eab\u88fd\u5099"},"content":{"rendered":"<!-- wp:paragraph -->\n<p>\u4f5c\u8005\u96f7\u5fb7\u548c\u91d1\u7d0d\u88e1\u00b7\u6c83\u68ee<\/p>\n<!-- \/wp:paragraph -->\n\n<!-- wp:paragraph -->\n<p>\u6458\u8981Swift 2S Turbo DNA\u5eab\u5957\u4ef6\u662f\u5b8c\u5168\u81ea\u52d5\u5316\u7684\uff0c\u4f7f\u7528\u4e2d\u5fc3OT-2\u5feb\u901f\uff0c\u653e\u624b\uff0c\u9ad8\u70baIllumina\u4e0b\u4e00\u4ee3\u5b9a\u5e8f\u63d0\u4f9b\u7684\u9ad8\u54c1\u8cea\u6587\u5eab\u3002\u4e3b\u8981\u767c\u73fe.8\u500b\u5e8f\u5217\u6e96\u5099\u7684DNA\u6587\u5eab\u53ef\u4ee5\u57283\u5c0f\u6642\u5167\u5efa\u69cb\u3002 .\u5206\u6790\u4e86326bp\u548c330bp\u7684\u76ee\u6a19\u63d2\u5165\u7269\u5927\u5c0f\u4eba\u985e\u57fa\u56e0\u7d44DNA\u3002 .\u4f4ePCR\u504f\u501a\uff0c\u986f\u793a\u5fae\u751f\u7269\u548c\u4eba\u985e\u6587\u5eab\u7684PCR\u91cd\u8907\u6c34\u6e96\u70ba\u5206\u5225\u70ba0.02%\u548c0.01%\u3002 .\u5728OT-2\u904b\u884c\u904e\u7a0b\u4e2d\u4fdd\u6301\u9ad8DNA\u8986\u84cb\u7387\uff0c\u900f\u904e\u76f8\u4f3c\u7684reads\u767e\u5206\u6bd4\u4f86\u63cf\u8ff0\u5c0d\u9f4a\uff1a\u5fae\u751f\u7269\u6587\u5eab\u4f5495.4%\uff0c\u4eba\u985e\u6587\u5eab\u4f5496.4%\u3002 .\u53ef\u9760\u4e14\u7cbe\u78ba\u7684\u6027\u80fd\u8996\u5b50\u71b1\u5faa\u74b0\u5668\u6a21\u7d44\u8207\u7b2c\u4e09\u65b9\u71b1\u5faa\u74b0\u5668\u76f8\u6bd4\uff0c\u900f\u904e\u76f8\u4f3c\u7684\u7522\u7387\u70ba5.1ng\/\u00b5land 7.36ng\/\u00b5l\uff0cCVs\u5206\u5225\u70ba13%\u548c14%\u3002<\/p>\n<!-- \/wp:paragraph -->\n\n<!-- wp:paragraph -->\n<p>\u4ecb\u7d39\u6b21\u4e16\u4ee3\u5b9a\u5e8f\uff08NGS\uff09\u6587\u5eab\u88fd\u5099\u662fRNA\u6216DNA\u7247\u6bb5\u7684\u904e\u7a0b\u9176\u6cd5\u5b9a\u5e8f\u7528\u7684\u4f7f\u7528NGS\u5eab\u7684\u6e96\u5099\u6b63\u5728\u8fc5\u901f\u589e\u52a0\uff0c\u70ba\u91ab\u7642\u4fdd\u5065\u548c\u751f\u547d\u79d1\u5b78\u7684\u7814\u7a76\u4eba\u54e1\u63d0\u4f9b\u4e86\u5404\u7a2e\u5de5\u4f5c\u6d41\u7a0b\uff0c\u4ee5\u5be6\u73fe\u66f4\u5feb\u3001\u66f4\u5bb9\u6613\u548c\u66f4\u9ad8\u7684\u6210\u672c\u5206\u6790\u548c\u89e3\u91cb\u5927\u578b\u57fa\u56e0\u7d44\u8cc7\u6599\u96c6\u7684\u6709\u6548\u65b9\u6cd5(1)\u3002 Swift 2S\u6e26\u8f2aDNA\u6587\u5eab\u6e96\u5099<\/p>\n<!-- \/wp:paragraph -->\n\n<!-- wp:paragraph -->\n<p>\u652f\u63f4\u5404\u7a2e\u6a23\u54c1\u7684\u6587\u5eab\u6e96\u5099\u5206\u5b50\u61c9\u7528\u3002\u9019\u500b\u81ea\u52d5\u5316\u53cb\u5584\u7684\u5de5\u4f5c\u6d41\u7a0b\u662f\u58d3\u7e2e\u7684\uff0c\u4f46\u8981\u624b\u52d5\u5b8c\u6210\u5de5\u4f5c\u6d41\u7a0b\u4ecd\u7136\u6703\u51fa\u73fe\u79fb\u6db2\u932f\u8aa4\uff0c\u9700\u8981\u5e7e\u500b\u932f\u8aa4\u55ce\u5e7e\u500b\u5c0f\u6642\u5b8c\u6210\u3002\u5728\u9019\u88e1\uff0c\u6211\u5011\u8aaa\u660e\u4e86Swift 2S\u6e26\u8f2a\u6587\u5eab\u88fd\u5099\u5fae\u751f\u7269\u7684\u5b8c\u5168\u81ea\u52d5\u5316\u4ee5\u53ca\u4eba\u985e\u57fa\u56e0\u7d44DNA\u3002\u8907\u96dc\u7684\u5eab\u88fd\u5099\u53ef\u4ee5\u5b8c\u5168\u81ea\u52d5\u5316\u7684\u8996\u4e2d\u5fc3OT- 2\uff0c\u6a21\u7d44\uff0c\u5be6\u9a57\u5ba4\u8edf\u9ad4\uff0c\u548c\u6b50\u7c73\u8304\u751f\u7269-tek\u6a19\u8a18\u7d50\u5408\u00ae\u7d14NGS\u73e0\u5b50\u3002<\/p>\n<!-- \/wp:paragraph -->\n\n<!-- wp:paragraph -->\n<p>\u6750\u6599\u8207\u65b9\u6cd5\u88fd\u5099\u4e86350bp\u63d2\u5165\u5927\u5c0f\u7684\u6587\u5eab~560bp\u6587\u5eab\u4f7f\u7528\u4eba\u985e\u57fa\u56e0\u7d44DNA(\u79d1\u91cc\u6c0f\u83cc. NA12878)\u548cEcoli\uff08ER2925\uff09gDNA\u7684GC\u542b\u91cf\u5206\u5225\u70ba40\u548c50%\u3002\u6bcf\u500bopentronOT-2\u904b\u884c\u5305\u542b7\u500b\u91cd\u8907\uff0c\u6bcf\u500b\u904b\u884c\u5305\u542b5\u621610\u500bPCR\u9031\u671f\uff0c\u53d6\u6c7a\u65bc\u555f\u52d5\u8f38\u5165\u3002\u4f7f\u7528\u7684\u7d50\u7d2e\u6bcd\u6df7\u5408\u5305\u62ec1\uff1a10\u7a00\u91cb\u7684\u8a66\u5291W4\u4ee5\u6e1b\u5c11\u9069\u914d\u5668\u4e8c\u805a\u9ad4\u3002\u6bcf\u500b\u6587\u5eab\u90fd\u5728Illumina MiSeq Nano v2 2x250\u8a66\u5291\u76d2\u4e0a\u9032\u884c\u5b9a\u5e8f\u3002\u8996\u8f49\u5b50\u71b1\u5faa\u74b0\u5668\u6a21\u7d44\u548c\u6eab\u5ea6\u6a21\u7d44\u7528\u65bc\u4e3b\u52d5\u51b7\u537b\u4e3b\u6df7\u5408\uff0c\u7d22\u5f15\u548c\u6a23\u672c\u4f7f\u7528Omega-Beo-tekMag-Bind\u00ae\u9032\u884c\u57fa\u65bc\u73e0\u5b50\u7684\u6e05\u7406\u7d14NGS\u73e0\u5b50(2)\u3002\u9019\u4e9b\u6a21\u7d44\u5171\u540c\u5141\u8a31\u9019\u500b\u8907\u5408\u5de5\u4f5c\u6d41\u7a0b\u7684\u5b8c\u5168\u81ea\u52d5\u5316\uff08\u57161\uff09\u3002 OT-2\u4f7f\u7528P300GEN2 8\u901a\u9053\u79fb\u6db2\u7ba1\u548cP50 GEN2\u55ae\u901a\u9053\u79fb\u6db2\u7ba1\u9032\u884c\u904b\u884cE.\u5927\u8178\u687f\u83ccgDNA\uff0cP300GEN2 8\u901a\u9053\u79fb\u6db2\u7ba1\u548cP20 GEN2\u55ae\u901a\u9053\u79fb\u6db2\u7ba1\u8207\u4eba\u985egDNA\u4e00\u8d77\u904b\u4f5c\u3002\u7136\u800c\uff0c\u6211\u5011\u5efa\u8b70P20. GEN2\u55ae\u901a\u9053\u79fb\u6db2\u7ba1\u6bcf\u6b21\u904b\u884c\uff0c\u4f7f\u75283\u500b\u5c16\u7aef\uff08\u57161\uff09\u3002\u7e3d\u904b\u884c\u6642\u9593\uff0c\u5305\u62ec\u52d5\u624b\u6642\u9593\u548c\u6a5f\u5668\u4eba\u904b\u884c\u6642\u9593\uff0c\u7d043\u5c0f\u6642\uff08\u57162\uff09\u3002\u5cf0\u503c\u7247\u6bb5\u5927\u5c0f\u4f7f\u7528\u5b89\u6377\u502b2100\u751f\u7269\u5206\u6790\u5100fastq\u6a94\u78ba\u5b9a.<\/p>\n<!-- \/wp:paragraph -->\n\n<!-- wp:paragraph -->\n<p>\u4f7f\u7528\u661f\u7cfb(7)\u4e2d\u7684FASTQC V0.11.9 (3)\u3001BOWTIE2 (4)\u3001Qualimap- BAMQC (5)\u548cBam\u8986\u84cbV2.4.1.0 (6)\u9032\u884c\u5206\u6790\uff0c\u4ee5\u78ba\u5b9a\u8986\u84cb\u7387\u3001\u63d2\u5165\u5927\u5c0f\u3001\u8b80\u53d6\u8207\u57fa\u56e0\u7d44\u5c0d\u9f4a\u3001GC\u542b\u91cf\u548c\u91cd\u8907\u767e\u5206\u6bd4\u3002 BWA\u5c0d\u9f4a\u5668V1.1.4\u9032\u884c\u9032\u4e00\u6b65\u5206\u6790\u3002<\/p>\n<!-- \/wp:paragraph -->\n\n<!-- wp:paragraph -->\n<p>\u7d50\u679c\u6587\u5eab\u7684\u7522\u91cf\u5177\u6709\u5f88\u9ad8\u7684\u53ef\u91cd\u8907\u6027\u548c\u53ef\u6bd4\u6027\u5728\u540c\u4e00\u904b\u884c\u7bc4\u570d\u5167\u7684\u8de8\u6a23\u672c\u548c\u8de8\u7269\u7a2e\u3002 Ecoli gDNA\u8f38\u5165100 ng\uff0c\u4eba\u985egDNA\u8f38\u516577 ng\u3002 . . Ecoli\u6587\u5eab\u7684\u5de5\u4f5c\u6d41\u7a0b\u5305\u62ec10\u500bPCR\u5faa\u74b0\uff0c\u5e73\u5747\u6fc3\u5ea6\u70ba20.34ng\/\u00b5l7%\u7684\u8b8a\u7570\u4fc2\u6578\uff08CV\uff09\u3002\u4eba\u985egDNA\u6587\u5eab\u5de5\u4f5c\u6d41\u7a0b\u5305\u62ec5\u500bPCR\u9031\u671f\uff0c\u5e73\u5747\u9031\u671f\u70ba. \u8a72\u6587\u5eab\u5177\u6709\u53ef\u6bd4\u6027\uff0c\u5e73\u5747\u5cf0\u7247\u6bb5\u5927\u5c0f\u70ba591bp\uff0cCV\u70ba7%. \u4eba\u985e\u6587\u5eab\u7684\u5e73\u5747\u5cf0\u503c\u7247\u6bb5\u5927\u5c0f\u70ba569bp\uff0cCV\u70ba4%\u3002\u7522\u91cf\u548c\u5cf0\u503c\u7247\u6bb5\u5927\u5c0fCV\u986f\u793a\u4e86\u4e0d\u540c\u4e95\u548c\u4e0d\u540c\u7269\u7a2e\u4e4b\u9593\u7684\u4e00\u81f4\u6027\uff08\u57163\uff09\u3002\u5b9a\u5e8f\u6307\u6a19\u5728\u4e0d\u540c\u6a23\u672c\u548c\u4e0d\u540c\u7269\u7a2e\u4e4b\u9593\u4e5f\u5177\u6709\u53ef\u6bd4\u6027\u3002\u5927\u8178\u687f\u83cc\u548c\u4eba\u985e\u6587\u5eab\u7684\u5e73\u5747\u63d2\u5165\u7269\u5927\u5c0f\u5206\u5225\u70ba326\u548c330 bp\uff0c\u8207\u4f9d\u8a66\u5291\u898f\u683c\u7d0d\u5165\u76f4\u63a5\u5b9a\u5e8f\u7684DNA\u76ee\u6a19\u9577\u5ea6\u76f8\u5c0d\u61c9\u3002\u5169\u500b\u5be6\u9a57\u90fd\u986f\u793a\u4e86\u7d0495%\u7684\u8b80\u53d6\u6620\u5c04\u6642\u8207\u53c3\u8003\u57fa\u56e0\u7d44\u5e8f\u5217\u5c0d\u9f4a\uff0c\u9ad8DNA\u56de\u6536\u7387\u548c\u5927\u5c0f\u9078\u64c7\u59cb\u7d42\u4fdd\u7559\u6bcf\u6b21\u904b\u884c\uff0c\u4ee5\u975e\u5e38\u4f4e\u7684PCR\u91cd\u8907\u4f86\u8aaa\u660e\uff08\u57164\uff09\u3002 . \u5728Ecoli\u6a23\u672c\uff08\u5716S1\uff09\u548c\u4eba\u985e\u6a23\u672c\uff08\u5716S2\uff09\u4e2d\u7684\u4e00\u81f4\u8986\u84cb\u986f\u793a\u4e86\u76f8\u540c\u7684\u6027\u80fd\u548c\u5e8f\u5217\u8868\u793a\u3002\u800c\u4e14\u55ae\u6838\u82f7\u9178\u8b8a\u7570\uff08SNV\uff09rs6061194\u70ba\u9019\u9032\u4e00\u6b65\u5c55\u793a\u4e86\u6027\u80fd\uff08\u5716S3\uff09\u3002\u6700\u5f8c\uff0c\u5c0d\u4e2d\u5fc3\u71b1\u5faa\u74b0\u5668\u6a21\u7d44\u7684\u6027\u80fd\u985e\u4f3c\u65bc\u624b\u52d5\u7b2c\u4e09\u65b9\u71b1\u5faa\u74b0\u5668(8)\uff0c\u5728\u7522\u91cf\u548cCV\u4e0a\u5177\u6709\u5171\u901a\u6027\u3002\u7522\u7387\u5206\u5225\u70ba5.1ng\/\u00b5land\u548c7.36ng\/\u00b5land\u8a72\u7aef\u5c0d\u7aefOpentrons\u5eab\u88fd\u5099\u7cfb\u7d71\u7684\u7cbe\u5ea6\u548c\u53ef\u9760\u6027\uff08\u88681\uff09\u3002<\/p>\n<!-- \/wp:paragraph -->\n\n<!-- wp:paragraph -->\n<p>\u7d50\u8ad6Opentrons OT-2\u53ef\u4ee5\u4e00\u6b21\u63d0\u4f9b8\u500b\u6216\u66f4\u591a\u7684\u9ad8\u54c1\u8cea\u5eab\uff0c\u81f3\u5c11\u4e0d\u52303\u500b\u5c0f\u6642\uff0c\u6700\u7d42\u6e1b\u5c11\u4e86\u624b\u52d5\u79fb\u6db2\u932f\u8aa4\u7684\u98a8\u96aa\uff0c\u4e26\u7bc0\u7701\u4e86\u7528\u6236\u6642\u9593\u3002 .\u53ef\u91cd\u8907\u7684\u548c\u4e00\u81f4\u7684\u6e2c\u5e8f\u6307\u6a19\u5728\u5fae\u751f\u7269\u548c\u4eba\u985e\u6587\u5eab\u7684\u7522\u91cf\u3001CV\u3001\u63d2\u5165\u5927\u5c0f\u3001\u5c0d\u9f4a\u3001PCR\u91cd\u8907\u548c\u8986\u84cb\u65b9\u9762\u3002 .\u7576\u7522\u7387\u548cCV\u70ba\u6642\uff0c\u8996\u4e2d\u5fc3\u71b1\u5faa\u74b0\u5668\u6a21\u7d44\u7684\u53ef\u9760\u548c\u7cbe\u78ba\u7684\u6027\u80fd\u8207\u7b2c\u4e09\u65b9\u71b1\u5faa\u74b0\u5668\u76f8\u6bd4\u3002<\/p>\n<!-- \/wp:paragraph -->\n\n<!-- wp:image {\"id\":3064,\"sizeSlug\":\"full\",\"linkDestination\":\"none\"} -->\n<figure class=\"wp-block-image size-full\"><img src=\"https:\/\/opentrons.com.cn\/wp-content\/uploads\/2024\/06\/image-115.png\" alt=\"\u9176\u548c\u53cc\u6307\u6807\u5728\u6e29\u5ea6\u6a21\u5757\u4e0a\u79ef\u6781\u51b7\u5374\" class=\"wp-image-3064\"><figcaption class=\"wp-element-caption\">\u57161\uff1a<em>\u9175\u7d20\u548c\u96d9\u6307\u6a19\u5728\u6eab\u5ea6\u6a21\u7d44\u4e0a\u7a4d\u6975\u51b7\u537b<\/em><\/figcaption><\/figure>\n<!-- \/wp:image -->\n\n<!-- wp:image {\"id\":3066,\"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-117.png\" alt=\"\" class=\"wp-image-3066\"><\/figure>\n<!-- \/wp:image -->\n\n<!-- wp:paragraph -->\n<p>\u57161. 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