{"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:\/\/opentrons.com.cn\/knowledge2\/xlzbqzdwswhrldnambdzb\/","title":{"rendered":"\u5e8f\u5217\u51c6\u5907\u5168\u81ea\u52a8\u5fae\u751f\u7269\u548c\u4eba\u7c7bDNA\u6a21\u677f\u7684\u6587\u5e93\u5236\u5907"},"content":{"rendered":"\n<p>\u4f5c\u8005<br>\u96f7\u5fb7\u548c\u91d1\u7eb3\u91cc\u00b7\u6c83\u68ee<\/p>\n\n\n\n<p>\u6458\u8981<br>Swift 2S Turbo DNA\u5e93\u5957\u4ef6\u662f\u5b8c\u5168\u81ea\u52a8\u5316\u7684\uff0c\u4f7f\u7528\u4e2d\u5fc3OT-2\u5feb\u901f\uff0c\u653e\u624b\uff0c\u9ad8\u4e3aIllumina\u4e0b\u4e00\u4ee3\u6d4b\u5e8f\u63d0\u4f9b\u7684\u9ad8\u8d28\u91cf\u6587\u5e93\u3002<br>\u4e3b\u8981\u53d1\u73b0<br>.8\u4e2a\u5e8f\u5217\u51c6\u5907\u7684DNA\u6587\u5e93\u53ef\u4ee5\u57283\u5c0f\u65f6\u5185\u6784\u5efa\u3002<br>.\u5206\u6790\u4e86326bp\u548c330bp\u7684\u76ee\u6807\u63d2\u5165\u7269\u5927\u5c0f\u4eba\u7c7b\u57fa\u56e0\u7ec4DNA\u3002<br>.\u4f4ePCR\u504f\u501a\uff0c\u8bf4\u660e\u5fae\u751f\u7269\u548c\u4eba\u7c7b\u6587\u5e93\u7684PCR\u91cd\u590d\u6c34\u5e73\u4e3a\u5206\u522b\u4e3a0.02%\u548c0.01%\u3002<br>.\u5728OT-2\u8fd0\u884c\u8fc7\u7a0b\u4e2d\u4fdd\u6301\u9ad8DNA\u8986\u76d6\u7387\uff0c\u901a\u8fc7\u76f8\u4f3c\u7684reads\u767e\u5206\u6bd4\u6765\u63cf\u8ff0\u5bf9\u9f50\uff1a\u5fae\u751f\u7269\u6587\u5e93\u536095.4%\uff0c\u4eba\u7c7b\u6587\u5e93\u536096.4%\u3002<br>.\u53ef\u9760\u548c\u7cbe\u786e\u7684\u6027\u80fd\u89c6\u5b50\u70ed\u5faa\u73af\u5668\u6a21\u5757\u4e0e\u7b2c\u4e09\u65b9\u70ed\u5faa\u73af\u5668\u76f8\u6bd4\uff0c\u901a\u8fc7\u76f8\u4f3c\u7684\u4ea7\u7387\u4e3a5.1ng\/\u00b5land 7.36ng\/\u00b5l\uff0cCVs\u5206\u522b\u4e3a13%\u548c14%\u3002<\/p>\n\n\n\n<p>\u4ecb\u7ecd<br>\u4e0b\u4e00\u4ee3\u6d4b\u5e8f\uff08NGS\uff09\u6587\u5e93\u5236\u5907\u662fRNA\u6216DNA\u7247\u6bb5\u7684\u8fc7\u7a0b\u9176\u6cd5\u6d4b\u5e8f\u7528\u7684\u4f7f\u7528NGS\u5e93\u7684\u51c6\u5907\u6b63\u5728\u8fc5\u901f\u589e\u52a0\uff0c\u4e3a\u533b\u7597\u4fdd\u5065\u548c\u751f\u547d\u79d1\u5b66\u7684\u7814\u7a76\u4eba\u5458\u63d0\u4f9b\u4e86\u5404\u79cd\u5de5\u4f5c\u6d41\u7a0b\uff0c\u4ee5\u5b9e\u73b0\u66f4\u5feb\u3001\u66f4\u5bb9\u6613\u548c\u66f4\u9ad8\u7684\u6210\u672c\u5206\u6790\u548c\u89e3\u91ca\u5927\u578b\u57fa\u56e0\u7ec4\u6570\u636e\u96c6\u7684\u6709\u6548\u65b9\u6cd5(1)\u3002Swift 2S\u6da1\u8f6eDNA\u6587\u5e93\u51c6\u5907<\/p>\n\n\n\n<p>\u652f\u6301\u5bf9\u5404\u79cd\u6837\u54c1\u7684\u6587\u5e93\u51c6\u5907<br>\u5206\u5b50\u5e94\u7528\u3002\u8fd9\u4e2a\u81ea\u52a8\u5316\u53cb\u597d\u7684\u5de5\u4f5c\u6d41\u662f\u538b\u7f29\u7684\uff0c\u4f46\u8981\u624b\u52a8\u5b8c\u6210\u5de5\u4f5c\u6d41\u4ecd\u7136\u4f1a\u51fa\u73b0\u79fb\u6db2\u9519\u8bef\uff0c\u9700\u8981\u51e0\u4e2a\u9519\u8bef\u5417\u51e0\u4e2a\u5c0f\u65f6\u5b8c\u6210\u3002\u5728\u8fd9\u91cc\uff0c\u6211\u4eec\u8bf4\u660e\u4e86Swift 2S\u6da1\u8f6e\u6587\u5e93\u5236\u5907\u5fae\u751f\u7269\u7684\u5b8c\u5168\u81ea\u52a8\u5316\u4ee5\u53ca\u4eba\u7c7b\u57fa\u56e0\u7ec4DNA\u3002\u590d\u6742\u7684\u5e93\u5236\u5907\u53ef\u4ee5\u5b8c\u5168\u81ea\u52a8\u5316\u7684\u89c6\u4e2d\u5fc3OT- 2\uff0c\u6a21\u5757\uff0c\u5b9e\u9a8c\u5ba4\u8f6f\u4ef6\uff0c\u548c\u6b27\u7c73\u8304\u751f\u7269-tek\u6807\u8bb0\u7ed3\u5408\u00ae\u7eafNGS\u73e0\u5b50\u3002<\/p>\n\n\n\n<p>\u6750\u6599\u548c\u65b9\u6cd5<br>\u5236\u5907\u4e86350bp\u63d2\u5165\u5927\u5c0f\u7684\u6587\u5e93~560bp\u6587\u5e93\u4f7f\u7528\u4eba\u7c7b\u57fa\u56e0\u7ec4DNA(\u79d1\u91cc\u6c0f\u83cc. NA12878)\u548cEcoli\uff08ER2925\uff09gDNA\u7684GC\u542b\u91cf\u5206\u522b\u4e3a40\u548c50%\u3002\u6bcf\u4e2aopentronOT-2\u8fd0\u884c\u5305\u542b7\u4e2a\u91cd\u590d\uff0c\u6bcf\u4e2a\u8fd0\u884c\u5305\u542b5\u4e2a\u621610\u4e2aPCR\u5468\u671f\uff0c\u53d6\u51b3\u4e8e\u542f\u52a8\u8f93\u5165\u3002\u4f7f\u7528\u7684\u7ed3\u624e\u6bcd\u6df7\u5408\u5305\u62ec1\uff1a10\u7a00\u91ca\u7684\u8bd5\u5242W4\u4ee5\u51cf\u5c11\u9002\u914d\u5668\u4e8c\u805a\u4f53\u3002\u6bcf\u4e2a\u6587\u5e93\u90fd\u5728Illumina MiSeq Nano v2 2&#215;250\u8bd5\u5242\u76d2\u4e0a\u8fdb\u884c\u6d4b\u5e8f\u3002\u89c6\u8f6c\u5b50\u70ed\u5faa\u73af\u5668\u6a21\u5757\u548c\u6e29\u5ea6\u6a21\u5757\u7528\u4e8e\u4e3b\u52a8\u51b7\u5374\u4e3b\u6df7\u5408\uff0c\u7d22\u5f15\u548c\u6837\u672c\u4f7f\u7528Omega-Beo-tekMag-Bind\u00ae\u8fdb\u884c\u57fa\u4e8e\u73e0\u5b50\u7684\u6e05\u7406\u7eafNGS\u73e0\u5b50(2)\u3002\u8fd9\u4e9b\u6a21\u5757\u5171\u540c\u5141\u8bb8\u8fd9\u4e2a\u590d\u5408\u5de5\u4f5c\u6d41\u7684\u5b8c\u5168\u81ea\u52a8\u5316\uff08\u56fe1\uff09\u3002<br>OT-2\u4f7f\u7528P300GEN2 8\u901a\u9053\u79fb\u6db2\u7ba1\u548cP50 GEN2\u5355\u901a\u9053\u79fb\u6db2\u7ba1\u8fdb\u884c\u8fd0\u884cE.\u5927\u80a0\u6746\u83ccgDNA\uff0cP300GEN2 8\u901a\u9053\u79fb\u6db2\u7ba1\u548cP20 GEN2\u5355\u901a\u9053\u79fb\u6db2\u7ba1\u4e0e\u4eba\u7c7bgDNA\u4e00\u8d77\u8fd0\u884c\u3002\u7136\u800c\uff0c\u6211\u4eec\u63a8\u8350P20<br>. GEN2\u5355\u901a\u9053\u79fb\u6db2\u7ba1\u6bcf\u6b21\u8fd0\u884c\uff0c\u4f7f\u75283\u4e2a\u5c16\u7aef\uff08\u56fe1\uff09\u3002\u603b\u8fd0\u884c\u65f6\u95f4\uff0c\u5305\u62ec\u52a8\u624b\u65f6\u95f4\u548c\u673a\u5668\u4eba\u8fd0\u884c\u65f6\u95f4\uff0c\u7ea6\u4e3a3\u5c0f\u65f6\uff08\u56fe2\uff09\u3002\u5cf0\u503c\u7247\u6bb5\u5927\u5c0f\u4f7f\u7528\u5b89\u6377\u4f262100\u751f\u7269\u5206\u6790\u4eeafastq\u6587\u4ef6\u786e\u5b9a.<\/p>\n\n\n\n<p>\u4f7f\u7528\u661f\u7cfb(7)\u4e2d\u7684FASTQC V0.11.9 (3)\u3001BOWTIE2 (4)\u3001Qualimap- BAMQC (5)\u548cBam\u8986\u76d6V2.4.1.0 (6)\u8fdb\u884c\u5206\u6790\uff0c\u4ee5\u786e\u5b9a\u8986\u76d6\u7387\u3001\u63d2\u5165\u5927\u5c0f\u3001\u8bfb\u53d6\u4e0e\u57fa\u56e0\u7ec4\u5bf9\u9f50\u3001GC\u542b\u91cf\u548c\u91cd\u590d\u767e\u5206\u6bd4\u3002BWA\u5bf9\u9f50\u5668V1.1.4\u8fdb\u884c\u8fdb\u4e00\u6b65\u5206\u6790\u3002<\/p>\n\n\n\n<p>\u7ed3\u679c<br>\u6587\u5e93\u7684\u4ea7\u91cf\u5177\u6709\u5f88\u9ad8\u7684\u53ef\u91cd\u590d\u6027\u548c\u53ef\u6bd4\u6027<br>\u5728\u540c\u4e00\u8fd0\u884c\u8303\u56f4\u5185\u7684\u8de8\u6837\u672c\u548c\u8de8\u7269\u79cd\u3002Ecoli gDNA\u8f93\u5165100 ng\uff0c\u4ebagDNA\u8f93\u516577 ng\u3002. . 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srcset=\"https:\/\/opentrons.com.cn\/wp-content\/uploads\/2024\/06\/image-118.png 813w, https:\/\/opentrons.com.cn\/wp-content\/uploads\/2024\/06\/image-118-300x128.png 300w, https:\/\/opentrons.com.cn\/wp-content\/uploads\/2024\/06\/image-118-768x327.png 768w, https:\/\/opentrons.com.cn\/wp-content\/uploads\/2024\/06\/image-118-576x245.png 576w\" sizes=\"(max-width: 813px) 100vw, 813px\" \/><figcaption class=\"wp-element-caption\"><em>\u56fe2\u3002\u5de5\u4f5c\u6d41\u5206\u89e3\u603b\u6570\u30028\u4e2a\u6837\u54c1\u7684\u51c6\u5907\u65f6\u95f4\u7ea6\u4e3a20\u5206\u949f\uff0c\u673a\u5668\u4eba\u8fd0\u884c\u65f6\u95f4\u4e3a2\u5c0f\u65f637\u5206\u949f\uff0c16\u4e2a\u6837\u54c1\u4e3a2\u5c0f\u65f648\u5206\u949f\u3002\u8fd9\u4e2a\u5b8c\u5168\u81ea\u52a8\u5316\u7684\u5de5\u4f5c\u6d41\u7a0b\u5305\u62ec10\u5206\u949f\u7684\u7247\u6bb5\u5316\u548c5\u4e2aPCR\u5faa\u73af\u3002<\/em><\/figcaption><\/figure>\n\n\n\n<figure class=\"wp-block-image size-full\"><img decoding=\"async\" width=\"806\" height=\"374\" src=\"https:\/\/opentrons.com.cn\/wp-content\/uploads\/2024\/06\/image-119.png\" alt=\"\u8de8\u5e93\u7684\u53ef\u8861\u91cf\u7684\u4ea7\u91cf\u548c\u7247\u6bb5\u5927\u5c0f\" class=\"wp-image-3068\" srcset=\"https:\/\/opentrons.com.cn\/wp-content\/uploads\/2024\/06\/image-119.png 806w, https:\/\/opentrons.com.cn\/wp-content\/uploads\/2024\/06\/image-119-300x139.png 300w, https:\/\/opentrons.com.cn\/wp-content\/uploads\/2024\/06\/image-119-768x356.png 768w, https:\/\/opentrons.com.cn\/wp-content\/uploads\/2024\/06\/image-119-576x267.png 576w\" sizes=\"(max-width: 806px) 100vw, 806px\" \/><figcaption 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https:\/\/opentrons.com.cn\/wp-content\/uploads\/2024\/06\/image-120-768x551.png 768w, https:\/\/opentrons.com.cn\/wp-content\/uploads\/2024\/06\/image-120-576x414.png 576w\" sizes=\"(max-width: 773px) 100vw, 773px\" \/><figcaption class=\"wp-element-caption\"><em>\u56fe4.\u53ef\u6bd4\u8f83\u7684\u63d2\u5165\u5927\u5c0f\uff0c\u8bfb\u53d6\u6570\u5bf9\u9f50<\/em><\/figcaption><\/figure>\n\n\n\n<figure class=\"wp-block-image size-full\"><img decoding=\"async\" width=\"797\" height=\"175\" src=\"https:\/\/opentrons.com.cn\/wp-content\/uploads\/2024\/06\/image-121.png\" alt=\"\u540c\u6837\u53ef\u9760\u7684\u5236\u5907\u5728\u4eba\u7c7b\u548c\u5fae\u751f\u7269\u5e93\u4e0eOT-2\uff0c\u5728\u4e0d\u540c\u7684\u5b9e\u9a8c\u5ba4\u8fdb\u884c\" class=\"wp-image-3070\" srcset=\"https:\/\/opentrons.com.cn\/wp-content\/uploads\/2024\/06\/image-121.png 797w, https:\/\/opentrons.com.cn\/wp-content\/uploads\/2024\/06\/image-121-300x66.png 300w, https:\/\/opentrons.com.cn\/wp-content\/uploads\/2024\/06\/image-121-768x169.png 768w, https:\/\/opentrons.com.cn\/wp-content\/uploads\/2024\/06\/image-121-576x126.png 576w\" sizes=\"(max-width: 797px) 100vw, 797px\" \/><figcaption class=\"wp-element-caption\"><em>\u88681\u3002\u540c\u6837\u53ef\u9760\u7684\u5236\u5907\u5728\u4eba\u7c7b\u548c\u5fae\u751f\u7269\u5e93\u4e0eOT-2\uff0c\u5728\u4e0d\u540c\u7684\u5b9e\u9a8c\u5ba4\u8fdb\u884c\u3002\u4ea7\u7387\u4e3a5.1ng\/\u00b5land 7.36ng\/\u00b5land\uff0c\u5206\u522b\u4e3a13%\u548c14%\uff0c\u8bc1\u660e\u4e86\u5149\u7535\u4e8c\u6781\u7ba1\u70ed\u5faa\u73af\u5668\u7684\u7cbe\u5ea6\u548c\u53ef\u9760\u6027\u3002<\/em><\/figcaption><\/figure>\n","protected":false},"excerpt":{"rendered":"<p>\u4f5c\u8005\u96f7\u5fb7\u548c\u91d1\u7eb3\u91cc\u00b7\u6c83\u68ee \u6458\u8981Swift 2S Turbo DNA\u5e93\u5957\u4ef6\u662f\u5b8c\u5168\u81ea\u52a8\u5316\u7684\uff0c\u4f7f\u7528\u4e2d\u5fc3OT-2\u5feb\u901f\uff0c\u653e 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