{"id":153,"date":"2021-09-26T16:12:09","date_gmt":"2021-09-26T08:12:09","guid":{"rendered":"https:\/\/www.chunxunbio.com\/?page_id=153"},"modified":"2022-07-29T11:27:32","modified_gmt":"2022-07-29T03:27:32","slug":"10x-genomics-3-%e6%8a%80%e6%9c%af","status":"publish","type":"page","link":"https:\/\/www.chunxunbio.com\/?page_id=153","title":{"rendered":"\u5355\u7ec6\u80de\u8f6c\u5f55\u7ec4\u6d4b\u5e8f"},"content":{"rendered":"\n<p><\/p>\n\n\n\n<div class=\"wp-block-buttons\">\n<div class=\"wp-block-button is-style-fill\"><a class=\"wp-block-button__link has-white-color has-text-color\" href=\"https:\/\/www.chunxunbio.com\/?page_id=38\">CX Genomics 3&#8242; \u6280\u672f<\/a><\/div>\n\n\n\n<div class=\"wp-block-button\"><a class=\"wp-block-button__link has-background\" href=\"https:\/\/www.chunxunbio.com\/?page_id=153\">10X Genomics 3&#8242; \u6280\u672f<\/a><\/div>\n\n\n\n<div class=\"wp-block-button is-style-fill\"><a class=\"wp-block-button__link\" href=\"https:\/\/www.chunxunbio.com\/?page_id=935\">\u751f\u4fe1\u5206\u6790\u793a\u4f8b<\/a><\/div>\n<\/div>\n\n\n\n<hr class=\"wp-block-separator has-text-color has-background has-black-background-color has-black-color is-style-wide\"\/>\n\n\n\n<p><\/p>\n\n\n\n<p class=\"has-text-align-center has-black-color has-text-color\" style=\"font-size:28px\"><strong>10X Genomics 3&#8242; \u6280\u672f<\/strong><\/p>\n\n\n\n<h3>&nbsp;<strong>1\u3001\u6280\u672f\u4f18\u52bf<\/strong><\/h3>\n\n\n\n<p class=\"has-text-align-left has-text-color\">&nbsp; &nbsp; &nbsp; &nbsp; &nbsp;&nbsp;\u901a\u91cf\u9ad8\uff1a\u4e00\u6b21\u53ef\u4ee5\u540c\u65f6\u6d4b8\u4e2a\u6837\u672c\uff0c\u6bcf\u4e2a\u6837\u672c\u6700\u591a\u53ef\u4ee5\u68c0\u6d4b\u5230\u4e0a\u4e07\u4e2a\u7ec6\u80de\u3002<\/p>\n\n\n\n<p class=\"has-text-align-left has-text-color\">&nbsp; &nbsp; &nbsp; &nbsp; &nbsp;&nbsp;\u5468\u671f\u77ed\uff1a7\u5206\u949f\u5185\u5b8c\u6210\u4e0a\u4e07\u4e2a\u7ec6\u80de\u5c01\u88c5\uff0c\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\n\n\n<p class=\"has-text-align-left has-text-color\">&nbsp; &nbsp; &nbsp; &nbsp; &nbsp;&nbsp;\u5355\u7ec6\u80de\u7387\u9ad8\uff1a\u5355\u4e2aGEMs\u6355\u83b7\u5230\u591a\u4e2a\u7ec6\u80de\u7684\u6982\u7387\u6781\u4f4e\uff08\uff1c0.9%\/1000cells\uff09\u3002<\/p>\n\n\n\n<p class=\"has-text-align-left has-text-color\">&nbsp; &nbsp; &nbsp; &nbsp; &nbsp;&nbsp;\u6355\u83b7\u6548\u7387\u9ad8\uff1a\u7ec6\u80de\u6355\u83b7\u6548\u7387\u9ad8\u8fbe65%\u3002<\/p>\n\n\n\n<p class=\"has-text-align-left has-text-color\">&nbsp; &nbsp; &nbsp; &nbsp; &nbsp;&nbsp;\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\u80bf\u7624\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\n\n\n<h3><strong>2\u3001\u5b9e\u9a8c\u64cd\u4f5c\u65b9\u6cd5\u53ca\u6d41\u7a0b<\/strong><\/h3>\n\n\n\n<p class=\"has-text-align-left has-text-color\">\u2003\u2003\u5982\u56fe\u6240\u793a\uff0c10X Genomics\u7684Chromium\u7cfb\u7edf\u5229\u7528\u5fae\u6d41\u63a7\u7cfb\u7edf\u5c06\u542b\u6709Barcode\u7684\u51dd\u80f6\u73e0\uff08Gel Beads\uff09\u3001\u7ec6\u80de\u548c\u9176\u7684\u6df7\u5408\u7269\u3001\u6cb9\u4e09\u8005\u6df7\u5408\uff0c\u5f62\u6210GEMs\uff08\u6cb9\u5305\u6c34\u7684\u5fae\u4f53\u7cfb\uff09\u3002GEMs\u5f62\u6210\u540e\uff0c\u7ec6\u80de\u5728\u5176\u4e2d\u88c2\u89e3\uff0c\u91ca\u653e\u51famRNA\uff0c\u51dd\u80f6\u73e0\u81ea\u52a8\u6eb6\u89e3\u91ca\u653e\u5927\u91cfbarcode\u5e8f\u5217\uff0c\u5229\u7528PloyT\u5f15\u7269\u6355\u83b7\u6db2\u6ef4\u4e2d\u7684mRNA\u3002\u968f\u540emRNA\u9006\u8f6c\u5f55\u4ea7\u751f\u5e26\u6709Barcode\u548cUMI\u4fe1\u606f\u7684cDNA\uff0c\u6784\u5efa\u6807\u51c6\u6d4b\u5e8f\u6587\u5e93\u3002<\/p>\n\n\n\n<figure class=\"wp-block-image size-large\"><img loading=\"lazy\" width=\"960\" height=\"360\" src=\"https:\/\/www.chunxunbio.com\/wp-content\/uploads\/2021\/10\/sketch_10x.png\" alt=\"\" class=\"wp-image-177\" srcset=\"https:\/\/www.chunxunbio.com\/wp-content\/uploads\/2021\/10\/sketch_10x.png 960w, https:\/\/www.chunxunbio.com\/wp-content\/uploads\/2021\/10\/sketch_10x-300x113.png 300w, https:\/\/www.chunxunbio.com\/wp-content\/uploads\/2021\/10\/sketch_10x-768x288.png 768w\" sizes=\"(max-width: 960px) 100vw, 960px\" \/><\/figure>\n\n\n\n<p class=\"has-text-align-center has-black-color has-text-color\">&nbsp;<\/p>\n\n\n\n<h4><strong>2.1 \u5355\u7ec6\u80de\u60ac\u6db2\u5236\u5907<\/strong><\/h4>\n\n\n\n<p class=\"has-text-align-left has-text-color\">&nbsp; &nbsp; &nbsp; &nbsp; &nbsp;&nbsp;\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\u5728\u8fdb\u884c\u5355\u7ec6\u80deRNA-seq\u4e4b\u524d\uff0c\u53ef\u4ee5\u4f7f\u7528\u51b7\u51bb\u4fdd\u5b58\u6216\u7532\u9187\u56fa\u5b9a\u6765\u50a8\u5b58\u7ec6\u80de\u3002\u5bf9\u4e8e\u8fdb\u884c\u5355\u7ec6\u80de\u6d4b\u5e8f\u5b9e\u9a8c\u7684\u6837\u672c\uff0c\u8981\u6c42\u7ec6\u80de\u6d3b\u6027&gt;70%\uff0c \u7ec6\u80de\u6d53\u5ea6\u5728700-1200\u7ec6\u80de\/\u03bcl\uff0c\u7ec6\u80de\u5c3a\u5bf8\u6700\u5927\u4e0d\u8d85\u8fc730\u03bcm\u3002<\/p>\n\n\n\n<h4><strong>2.2 \u7ec6\u80de\u5206\u9009<\/strong><\/h4>\n\n\n\n<p class=\"has-text-align-left has-text-color\">&nbsp; &nbsp; &nbsp; &nbsp; &nbsp;&nbsp;\u5982\u4e0a\u56fe\u6240\u793a\uff0c10X Genomics\u8fd0\u7528\u5fae\u6d41\u63a7\u7cfb\u7edf\u8fdb\u884c\u7ec6\u80de\u5206\u9009\uff0c\u51dd\u80f6\u73e0\u5300\u901f\u5730\u4ece\u5de6\u65b9\uff08\u56fe\u793a\uff09\u8fdb\u5165\uff0c\u5f85\u5206\u9009\u7684\u7ec6\u80de\u548c\u9176\u4ece\u4e0b\u65b9\uff08\u56fe\u793a\uff09\u4ee5\u4e00\u5b9a\u65f6\u95f4\u95f4\u9694\u8fdb\u5165\uff0c\u4e0e\u51dd\u80f6\u73e0\u4e00\u8d77\u6cb9\u76f8\u4e2d\u5f62\u6210GEMs\uff08Gel Bead in emulsion\uff09\u3002\u5982\u56fe\u6240\u793a\uff0c\u51dd\u80f6\u73e0\u4e0a\u7684\u5be1\u6838\u82f7\u9178\u94fe\u4f9d\u6b21\u4e3aIllumina TruSeq Read 1\u6d4b\u5e8f\u5f15\u7269\u300116bp 10X Barcode\u5e8f\u5217\uff08\u7528\u4e8e\u533a\u5206\u7ec6\u80de\uff09\u300112 bp unique molecular identifier\uff08UMI\uff0c\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\u53cd\u8f6c\u5f55\u5f15\u7269\u3002\u5728\u968f\u540e\u7684\u6587\u5e93\u6784\u5efa\u4e2d\uff0c\u76f8\u540c\u7684\u7ec6\u80de\u6269\u589e\u51fa\u7684cDNA\u5e26\u6709\u76f8\u540c\u768410x Barcode\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\u4e2aGEMs\u53ea\u4f1a\u5305\u542b\u4e00\u4e2a\u7ec6\u80de\u548c\u4e00\u4e2a\u51dd\u80f6\u73e0\uff0c\u4f46\u6709\u65f6\u5019\u4e5f\u4f1a\u51fa\u73b0\u4e00\u4e2aGEM\u91cc\u6ca1\u6709\u6216\u6709\u591a\u4e2a\u7ec6\u80de\u7684\u60c5\u51b5\uff0c\u5bf9\u4e8e\u8fd9\u79cdGEMs\u5219\u9700\u8981\u901a\u8fc7\u5728\u540e\u7eed\u5206\u6790\u4e2d\u8bc6\u522b\u51fa\u6765\u5e76\u8fdb\u884c\u53bb\u9664\u3002<\/p>\n\n\n\n<div class=\"wp-block-image\"><figure class=\"aligncenter size-large\"><img loading=\"lazy\" width=\"500\" height=\"280\" src=\"https:\/\/www.chunxunbio.com\/wp-content\/uploads\/2021\/10\/bead_10x.png\" alt=\"\" class=\"wp-image-165\" srcset=\"https:\/\/www.chunxunbio.com\/wp-content\/uploads\/2021\/10\/bead_10x.png 500w, https:\/\/www.chunxunbio.com\/wp-content\/uploads\/2021\/10\/bead_10x-300x168.png 300w\" sizes=\"(max-width: 500px) 100vw, 500px\" \/><\/figure><\/div>\n\n\n\n<p class=\"has-text-align-center has-black-color has-text-color\">&nbsp;<\/p>\n\n\n\n<h4><strong>2.3 cDNA\u751f\u6210\u4e0e\u6269\u589e<\/strong><\/h4>\n\n\n\n<p class=\"has-text-align-left has-text-color\">&nbsp; &nbsp; &nbsp; &nbsp; &nbsp;&nbsp;\u5f62\u6210GEMs\u540e\uff0c\u51dd\u80f6\u73e0\u6eb6\u89e3\uff0c\u7ec6\u80de\u88c2\u89e3\u91ca\u653emRNA\uff0c\u51dd\u80f6\u73e0\u4e2d\u7684polyT\u6355\u83b7mRNA\u4e2d\u7684polyA\uff0c\u5e76\u9006\u8f6c\u5f55\u751f\u6210\u7528\u4e8e\u6d4b\u5e8f\u7684cDNA\u3002\u7136\u540e\u7834\u574f\u6cb9\u5305\u6c34\u7ed3\u6784\uff0c\u4f7f\u7528\u78c1\u73e0\u7eaf\u5316cDNA\u5e76\u8fdb\u884c\u6e05\u6d17\uff0c\u518d\u8fdb\u884c\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\n\n\n<div class=\"wp-block-image\"><figure class=\"aligncenter size-large\"><img loading=\"lazy\" width=\"480\" height=\"440\" src=\"https:\/\/www.chunxunbio.com\/wp-content\/uploads\/2021\/10\/cdna_10x.png\" alt=\"\" class=\"wp-image-169\" srcset=\"https:\/\/www.chunxunbio.com\/wp-content\/uploads\/2021\/10\/cdna_10x.png 480w, https:\/\/www.chunxunbio.com\/wp-content\/uploads\/2021\/10\/cdna_10x-300x275.png 300w\" sizes=\"(max-width: 480px) 100vw, 480px\" \/><\/figure><\/div>\n\n\n\n<p class=\"has-text-align-center\">&nbsp;<\/p>\n\n\n\n<div class=\"wp-block-image\"><figure class=\"aligncenter size-large\"><img loading=\"lazy\" width=\"480\" height=\"220\" src=\"https:\/\/www.chunxunbio.com\/wp-content\/uploads\/2021\/10\/preamplify_10x.png\" alt=\"\" class=\"wp-image-173\" srcset=\"https:\/\/www.chunxunbio.com\/wp-content\/uploads\/2021\/10\/preamplify_10x.png 480w, https:\/\/www.chunxunbio.com\/wp-content\/uploads\/2021\/10\/preamplify_10x-300x138.png 300w\" sizes=\"(max-width: 480px) 100vw, 480px\" \/><\/figure><\/div>\n\n\n\n<p class=\"has-text-align-center has-black-color has-text-color\">&nbsp;<\/p>\n\n\n\n<h4><strong>2.4 \u6587\u5e93\u6784\u5efa<\/strong><\/h4>\n\n\n\n<p class=\"has-text-align-left has-text-color\">&nbsp; &nbsp; &nbsp; &nbsp; &nbsp;&nbsp;cDNA\u6269\u589e\u5b8c\u6210\u540e\uff0c\u5229\u7528\u9176\u5c06cDNA\u6253\u65ad\u6210\u7247\u6bb5\uff0c\u901a\u8fc7\u672b\u7aef\u4fee\u590d\u3001\u5e76\u52a0\u5165A\u78b1\u57fa\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\n\n\n<div class=\"wp-block-image\"><figure class=\"aligncenter size-large\"><img loading=\"lazy\" width=\"480\" height=\"440\" src=\"https:\/\/www.chunxunbio.com\/wp-content\/uploads\/2021\/10\/library_10x.png\" alt=\"\" class=\"wp-image-171\" srcset=\"https:\/\/www.chunxunbio.com\/wp-content\/uploads\/2021\/10\/library_10x.png 480w, https:\/\/www.chunxunbio.com\/wp-content\/uploads\/2021\/10\/library_10x-300x275.png 300w\" sizes=\"(max-width: 480px) 100vw, 480px\" \/><\/figure><\/div>\n\n\n\n<p class=\"has-text-align-center has-black-color has-text-color\">&nbsp;<\/p>\n\n\n\n<h4><strong>2.5 \u6587\u5e93\u6d4b\u5e8f<\/strong><\/h4>\n\n\n\n<p class=\"has-text-align-left has-text-color\">&nbsp; &nbsp; &nbsp; &nbsp; &nbsp;&nbsp;\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\n\n\n<div class=\"wp-block-image\"><figure class=\"aligncenter size-large\"><img loading=\"lazy\" width=\"750\" height=\"790\" src=\"https:\/\/www.chunxunbio.com\/wp-content\/uploads\/2021\/10\/sequence.png\" alt=\"\" class=\"wp-image-175\" srcset=\"https:\/\/www.chunxunbio.com\/wp-content\/uploads\/2021\/10\/sequence.png 750w, https:\/\/www.chunxunbio.com\/wp-content\/uploads\/2021\/10\/sequence-285x300.png 285w\" sizes=\"(max-width: 750px) 100vw, 750px\" \/><\/figure><\/div>\n\n\n\n<p class=\"has-text-align-center has-black-color has-text-color\">&nbsp;<\/p>\n","protected":false},"excerpt":{"rendered":"<p>10X Genomics 3&#8242; \u6280\u672f &nbsp;1\u3001\u6280\u672f\u4f18\u52bf &nbsp; &nbsp; &#038;nb [&hellip;]<\/p>\n","protected":false},"author":1,"featured_media":0,"parent":38,"menu_order":0,"comment_status":"closed","ping_status":"closed","template":"","meta":[],"_links":{"self":[{"href":"https:\/\/www.chunxunbio.com\/index.php?rest_route=\/wp\/v2\/pages\/153"}],"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=153"}],"version-history":[{"count":50,"href":"https:\/\/www.chunxunbio.com\/index.php?rest_route=\/wp\/v2\/pages\/153\/revisions"}],"predecessor-version":[{"id":1147,"href":"https:\/\/www.chunxunbio.com\/index.php?rest_route=\/wp\/v2\/pages\/153\/revisions\/1147"}],"up":[{"embeddable":true,"href":"https:\/\/www.chunxunbio.com\/index.php?rest_route=\/wp\/v2\/pages\/38"}],"wp:attachment":[{"href":"https:\/\/www.chunxunbio.com\/index.php?rest_route=%2Fwp%2Fv2%2Fmedia&parent=153"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}