{"id":38,"date":"2020-09-08T17:30:06","date_gmt":"2020-09-08T09:30:06","guid":{"rendered":"http:\/\/localhost\/PureCell\/?page_id=38"},"modified":"2022-08-24T15:35:18","modified_gmt":"2022-08-24T07:35:18","slug":"%e5%8d%95%e7%bb%86%e8%83%9e%e8%bd%ac%e5%bd%95%e7%bb%84%e6%b5%8b%e5%ba%8f","status":"publish","type":"page","link":"https:\/\/www.chunxunbio.com\/?page_id=38","title":{"rendered":"\u5355\u7ec6\u80de\u8f6c\u5f55\u7ec4\u6d4b\u5e8f"},"content":{"rendered":"\n
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CX Genomics 3′ \u6280\u672f<\/a><\/div>\n\n\n\n
10X Genomics 3′ \u6280\u672f<\/a><\/div>\n\n\n\n
\u751f\u4fe1\u5206\u6790\u793a\u4f8b<\/a><\/div>\n<\/div>\n\n\n

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CX Genomics 3′ \u6280\u672f<\/strong><\/p>\n

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1\u3001\u6280\u672f\u4f18\u52bf<\/strong><\/h3>\n

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\u00a0 \u00a0 \u00a0 \u00a0 \u00a0\u00a0\u9ad8\u7cbe\u5ea6\uff1a\u5355\u7ec6\u80de\u7cbe\u5ea6\u8fbe\u523090%\u4ee5\u4e0a<\/p>\n

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\u00a0 \u00a0 \u00a0 \u00a0 \u00a0\u00a0\u9ad8\u901a\u91cf\uff1a\u6bcf\u4e2a\u6837\u672c\u6700\u591a\u53ef\u4ee5\u68c0\u6d4b\u5230\u4e0a\u4e07\u4e2a\u7ec6\u80de<\/p>\n

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\u00a0 \u00a0 \u00a0 \u00a0 \u00a0\u00a0\u5468\u671f\u77ed\uff1a\u4e00\u5929\u4e4b\u5185\u5b8c\u6210\u7ec6\u80de\u60ac\u6db2\u5236\u5907\u3001\u5355\u7ec6\u80de\u6355\u83b7\u3001\u6269\u589e\u4ee5\u53ca\u5efa\u5e93\u3002<\/p>\n

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\u00a0 \u00a0 \u00a0 \u00a0 \u00a0\u00a0\u5e94\u7528\u8303\u56f4\u5e7f\uff1a\u52a8\u7269\u7ec6\u80de\u548c\u690d\u7269\u7ec6\u80de\u5747\u53ef\u4ee5\u8fdb\u884c\u5355\u7ec6\u80de\u6d4b\u5e8f\uff0c\u5df2\u5e7f\u6cdb\u5e94\u7528\u4e8e\u7ec6\u80de\u5f02\u8d28\u6027\u3001\u514d\u75ab\u7ec6\u80de\u7fa4\u4f53\u68c0\u6d4b\u548c\u80da\u80ce\u53d1\u80b2\u7814\u7a76\u7b49\u3002<\/p>\n

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2\u3001\u5b9e\u9a8c\u64cd\u4f5c\u65b9\u6cd5\u53ca\u6d41\u7a0b<\/strong><\/h3>\n

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\u2003\u2003\u5982\u56fe\u6240\u793a\uff0cPure-Cell\u5355\u7ec6\u80de\u6d4b\u5e8f\u7cfb\u7edf\u5229\u7528\u5fae\u6d41\u63a7\u7cfb\u7edf\u5c06\u542b\u6709Barcode\u3001UMI\uff08unique molecular identifier\uff09\u548cPoly\uff08dT\uff09\u7684Bead\u3001\u7ec6\u80de\u548c\u6cb9\u4e09\u8005\u6df7\u5408\uff0c\u5f62\u6210\u4e0a\u4e07\u4e2a\u7eb3\u7c73\u7ea7\u6cb9\u5305\u6c34\u7684\u5fae\u4f53\u7cfb\u3002\u6cb9\u5305\u6c34\u7ed3\u6784\u5f62\u6210\u540e\uff0c\u7ec6\u80de\u5728\u5176\u4e2d\u88c2\u89e3\uff0c\u91ca\u653e\u51famRNA\uff0cBead\u5229\u7528PloyT\u5f15\u7269\u6355\u83b7\u6db2\u6ef4\u4e2d\u7684mRNA\u3002\u968f\u540e\u7834\u574f\u6cb9\u5305\u6c34\u7ed3\u6784\uff0c\u6536\u83b7\u6355\u83b7mRNA\u7684Beads\uff0c\u8fdb\u884c\u9006\u8f6c\u5f55\uff0c\u4ea7\u751f\u5e26\u6709Barcode\u548cUMI\u4fe1\u606f\u7684cDNA\uff0c\u6784\u5efa\u6807\u51c6\u6d4b\u5e8f\u6587\u5e93\u3002<\/p>\n

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2.1 \u5355\u7ec6\u80de\u60ac\u6db2\u5236\u5907<\/strong><\/h4>\n

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\u2003\u2003\u65b0\u9c9c\u7ec4\u7ec7\u6837\u672c\u9700\u8981\u6d88\u5316\u6210\u5355\u7ec6\u80de\u60ac\u6db2\uff0c\u5bf9\u4e8e\u57f9\u517b\u7684\u7ec6\u80de\u6216\u5df2\u7ecf\u5904\u4e8e\u60ac\u6d6e\u72b6\u6001\u7684\u7ec6\u80de\uff0c\u9700\u8981\u5bf9\u7ec6\u80de\u6d17\u6da4\u4ee5\u53bb\u9664\u57f9\u517b\u57fa\u3002\u5355\u7ec6\u80de\u60ac\u6db2\u5236\u5907\u5b8c\u6210\u540e\u9700\u5c3d\u5feb\u8fdb\u884c\u4e0a\u673a\u6355\u83b7\uff0c\u4e5f\u53ef\u901a\u8fc7\u7ec6\u80de\u4fdd\u5b58\u6db2\u8fdb\u884c\u77ed\u65f6\u95f4\u5185\u4fdd\u5b58\u3002\u5355\u7ec6\u80de\u8f6c\u5f55\u7ec4\u6d4b\u5e8f\u8981\u6c42\u6837\u672c\u7ec6\u80de\u6d3b\u7387>85%\uff0c\u7ec6\u80de\u6d53\u5ea6\u5728700-1200\u7ec6\u80de\/\u03bcl\uff0c\u7ec6\u80de\u6570\u91cf\u57285\u00d7106\u4ee5\u4e0a\u3002\/p><\/p>\n

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2.2 \u7ec6\u80de\u5206\u9009<\/strong><\/h4>\n

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\u2003\u2003\u5982\u4e0a\u56fe\u6240\u793a\uff0cPure-Cell\u5355\u7ec6\u80de\u6d4b\u5e8f\u7cfb\u7edf\u8fd0\u7528\u5fae\u6d41\u63a7+\u4eba\u5de5\u667a\u80fd\u7cfb\u7edf\u8fdb\u884c\u7ec6\u80de\u5206\u9009\uff0c\u60ac\u6d6e\u5728\u88c2\u89e3\u6db2\u7684Beads\u5300\u901f\u5730\u4ece\u5de6\u65b9\uff08\u56fe\u793a\uff09\u8fdb\u5165\uff0c\u5f85\u5206\u9009\u7684\u7ec6\u80de\u4ece\u4e0b\u65b9\uff08\u56fe\u793a\uff09\u4ee5\u4e00\u5b9a\u65f6\u95f4\u95f4\u9694\u8fdb\u5165\uff0c\u4e0eBeads\u4e00\u8d77\u8fdb\u5165\u6cb9\u76f8\u4e2d\u5f62\u6210\u6cb9\u5305\u6c34\u7ed3\u6784\u3002\u5982\u4e0b\u56fe\u6240\u793a\uff0cBeads\u4e0a\u7684\u5be1\u6838\u82f7\u9178\u94fe\u4f9d\u6b21\u4e3aIllumina Nextera Read 1\u6d4b\u5e8f\u5f15\u7269\u300112bp Barcode\u5e8f\u5217\uff08\u7528\u4e8e\u533a\u5206\u7ec6\u80de\uff09\u30018bp UMI\uff08\u533a\u5206\u7ec6\u80de\u7684\u4e0d\u540c\u8f6c\u5f55\u672c\u5e76\u53bb\u9664PCR Duplications\uff0c\u964d\u4f4e\u6269\u589e\u504f\u597d\u6027\uff09\u300130 bp poly dT\u3002\u5728\u968f\u540e\u7684\u6587\u5e93\u6784\u5efa\u4e2d\uff0c\u76f8\u540c\u7684\u7ec6\u80de\u6269\u589e\u51fa\u7684cDNA\u5e26\u6709\u76f8\u540c\u7684\u7ec6\u80deBarcode\uff0c\u5176\u4e2d\u6765\u81ea\u76f8\u540c\u7684\u8f6c\u5f55\u672c\u7684cDNA\u4f1a\u5e26\u6709\u76f8\u540c\u7684UMI\uff0c\u7531\u6b64\uff0c\u4fbf\u53ef\u533a\u5206\u51fa\u4e0d\u540c\u7ec6\u80de\u6269\u589e\u51fa\u7684cDNA\u3002\u4e00\u822c\u800c\u8a00\uff0c\u6bcf\u4e2a\u6cb9\u5305\u6c34\u53ea\u4f1a\u5305\u542b\u4e00\u4e2a\u7ec6\u80de\u548c\u4e00\u4e2a\u51dd\u80f6\u73e0\uff0c\u4f46\u6709\u65f6\u5019\u4e5f\u4f1a\u51fa\u73b0\u5176\u4ed6\u60c5\u51b5\uff0c\u5bf9\u4e8e\u8fd9\u79cd\u60c5\u51b5\u5219\u9700\u8981\u901a\u8fc7\u5728\u540e\u7eed\u5206\u6790\u4e2d\u8bc6\u522b\u51fa\u6765\u5e76\u8fdb\u884c\u53bb\u9664\u3002<\/p>\n

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2.3 cDNA\u751f\u6210\u4e0e\u6269\u589e<\/strong><\/h4>\n

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\u2003\u2003\u5728\u6cb9\u5305\u6c34\u7ed3\u6784\u4e2d\uff0cBeads\u4e2d\u7684polyT\u6355\u83b7mRNA\u4e2d\u7684polyA\u3002\u968f\u540e\u8fdb\u884c\u7834\u6cb9\uff0c\u5e76\u8fdb\u884c\u9006\u8f6c\u5f55\u53cd\u5e94\u751f\u6210cDNA\u3002\u968f\u540e\u8fdb\u884ccDNA\u7684\u6269\u589e\u53cd\u5e94\uff0c\u7eaf\u5316\u540e\u8fdb\u884c\u8d28\u68c0\uff0c\u8d28\u68c0\u5408\u683c\u540e\u8fdb\u884c\u6587\u5e93\u6784\u5efa\u3002<\/p>\n

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2.4 \u6587\u5e93\u6784\u5efa<\/strong><\/h4>\n

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\u2003\u2003cDNA\u6269\u589e\u5b8c\u6210\u540e\uff0c\u5229\u7528Tn5\u9176\u5c06cDNA\u6253\u65ad\u6210\u7247\u6bb5\uff0c\u5e76\u52a0\u5165\u6d4b\u5e8f\u5f15\u7269\u63a5\u5934\uff0c\u8fde\u63a5Read2\u6d4b\u5e8f\u5f15\u7269\uff0c\u5e76\u901a\u8fc7PCR\u6269\u589e\u5f15\u5165\u6837\u54c1Index\u3001P5\u3001P7\u63a5\u5934\uff0c\u6700\u540e\u8fdb\u884c\u7247\u6bb5\u7b5b\u9009\uff0c\u4ece\u800c\u6784\u5efa\u542b\u6709P5\u548cP7\u63a5\u5934\u7684cDNA\u6587\u5e93\uff0c\u6587\u5e93\u8d28\u68c0\u5408\u683c\u540e\u518d\u4e0a\u673a\u6d4b\u5e8f\u3002<\/p>\n

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2.5 \u6587\u5e93\u6d4b\u5e8f<\/strong><\/h4>\n

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\u2003\u2003\u6587\u5e93\u8d28\u68c0\u5408\u683c\u540e\uff0c\u5c31\u4f7f\u7528Illumina\u53cc\u7aef\u6d4b\u5e8f\u5e73\u53f0\u8fdb\u884c\u6d4b\u5e8f\u3002\u5982\u56fe\u6240\u793a\uff0cIllumina\u6d4b\u5e8f\u7684\u6838\u5fc3\u5728\u4e8e\u5229\u7528\u53ef\u9006\u7ec8\u6b62\u7684\u3001\u8367\u5149\u6807\u8bb0\u7684dNTP\u8fdb\u884c\u8fb9\u5408\u6210\u8fb9\u6d4b\u5e8f\uff08Sequencing-By-Synthesis,SBS\uff09\u3002<\/p>\n

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<\/p>","protected":false},"excerpt":{"rendered":"

\u00a0 CX Genomics 3′ \u6280\u672f 1\u3001\u6280\u672f\u4f18\u52bf \u00a0 \u00a0 \u00a0 \u00a0 \u00a0\u00a0\u9ad8\u7cbe\u5ea6\uff1a\u5355\u7ec6\u80de\u7cbe\u5ea6\u8fbe\u52309 […]<\/p>\n","protected":false},"author":1,"featured_media":1005,"parent":34,"menu_order":2,"comment_status":"closed","ping_status":"closed","template":"","meta":[],"_links":{"self":[{"href":"https:\/\/www.chunxunbio.com\/index.php?rest_route=\/wp\/v2\/pages\/38"}],"collection":[{"href":"https:\/\/www.chunxunbio.com\/index.php?rest_route=\/wp\/v2\/pages"}],"about":[{"href":"https:\/\/www.chunxunbio.com\/index.php?rest_route=\/wp\/v2\/types\/page"}],"author":[{"embeddable":true,"href":"https:\/\/www.chunxunbio.com\/index.php?rest_route=\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"https:\/\/www.chunxunbio.com\/index.php?rest_route=%2Fwp%2Fv2%2Fcomments&post=38"}],"version-history":[{"count":41,"href":"https:\/\/www.chunxunbio.com\/index.php?rest_route=\/wp\/v2\/pages\/38\/revisions"}],"predecessor-version":[{"id":1179,"href":"https:\/\/www.chunxunbio.com\/index.php?rest_route=\/wp\/v2\/pages\/38\/revisions\/1179"}],"up":[{"embeddable":true,"href":"https:\/\/www.chunxunbio.com\/index.php?rest_route=\/wp\/v2\/pages\/34"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/www.chunxunbio.com\/index.php?rest_route=\/wp\/v2\/media\/1005"}],"wp:attachment":[{"href":"https:\/\/www.chunxunbio.com\/index.php?rest_route=%2Fwp%2Fv2%2Fmedia&parent=38"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}