{"id":381,"date":"2024-04-25T20:04:37","date_gmt":"2024-04-25T20:04:37","guid":{"rendered":"https:\/\/winterdev.cc\/nwgc-dev\/?page_id=381"},"modified":"2024-04-25T20:04:37","modified_gmt":"2024-04-25T20:04:37","slug":"library-prep","status":"publish","type":"page","link":"https:\/\/nwgc.gs.washington.edu\/wordpress\/services\/functional-genomics\/fiberseq\/library-prep\/","title":{"rendered":"Library Prep"},"content":{"rendered":"<p><a href=\"https:\/\/cdn.10xgenomics.com\/image\/upload\/v1666737555\/support-documents\/CG000338_ChromiumNextGEM_Multiome_ATAC_GEX_User_Guide_RevF.pdf\">Chromium Next GEM Single Cell Multiome ATAC + Gene Expression Library Prep Post-GEM Generation <\/a><\/p>\n<h2 class=\"wp-block-heading\">ATAC Library Construction<\/h2>\n<p>P7 and a sample index are added to pre-amplified transposed DNA during ATAC library construction via PCR. The final ATAC libraries contain the P5 and P7 sequences used in Illumina\u00ae bridge amplification.<\/p>\n<figure class=\"wp-block-image size-full is-resized\"><img loading=\"lazy\" decoding=\"async\" width=\"819\" height=\"494\" src=\"https:\/\/winterdev.cc\/nwgc-dev\/wp-content\/uploads\/2024\/08\/image-15.png\" alt=\"\" class=\"wp-image-1278\" style=\"width:630px;height:auto\" srcset=\"https:\/\/nwgc.gs.washington.edu\/wordpress\/wp-content\/uploads\/2024\/08\/image-15.png 819w, https:\/\/nwgc.gs.washington.edu\/wordpress\/wp-content\/uploads\/2024\/08\/image-15-300x181.png 300w, https:\/\/nwgc.gs.washington.edu\/wordpress\/wp-content\/uploads\/2024\/08\/image-15-768x463.png 768w, https:\/\/nwgc.gs.washington.edu\/wordpress\/wp-content\/uploads\/2024\/08\/image-15-375x226.png 375w\" sizes=\"auto, 100vw\" \/><\/figure>\n<h2 class=\"wp-block-heading\">Gene Expression Library Construction<\/h2>\n<p>Enzymatic fragmentation and size selection are used to optimize the cDNA amplicon size. P5, P7, i7 and i5 sample indexes, and TruSeq Read 2 (read 2 primer sequence) are added via End Repair, A-tailing, Adaptor Ligation, and PCR. The final gene expression libraries contain the P5 and P7 primers used in Illumina\u00ae bridge amplification.<\/p>\n<figure class=\"wp-block-image size-full is-resized\"><img loading=\"lazy\" decoding=\"async\" width=\"723\" height=\"658\" src=\"https:\/\/winterdev.cc\/nwgc-dev\/wp-content\/uploads\/2024\/08\/image-16.png\" alt=\"\" class=\"wp-image-1279\" style=\"width:588px;height:auto\" srcset=\"https:\/\/nwgc.gs.washington.edu\/wordpress\/wp-content\/uploads\/2024\/08\/image-16.png 723w, https:\/\/nwgc.gs.washington.edu\/wordpress\/wp-content\/uploads\/2024\/08\/image-16-300x273.png 300w, https:\/\/nwgc.gs.washington.edu\/wordpress\/wp-content\/uploads\/2024\/08\/image-16-375x341.png 375w\" sizes=\"auto, (max-width: 723px) 100vw, 723px\" \/><\/figure>\n","protected":false},"excerpt":{"rendered":"<p>Chromium Next GEM Single Cell Multiome ATAC + Gene Expression Library Prep Post-GEM Generation ATAC Library Construction P7 and a sample index are added to pre-amplified transposed DNA during ATAC library construction via PCR. The final ATAC libraries contain the P5 and P7 sequences used in Illumina\u00ae bridge amplification. Gene Expression Library Construction Enzymatic fragmentation and size selection are used to optimize the cDNA amplicon size. P5, P7, i7 and i5 sample indexes, and TruSeq Read 2 (read 2 primer&#8230;<\/p>\n<div><a class=\"more-link\" href=\"https:\/\/nwgc.gs.washington.edu\/wordpress\/services\/functional-genomics\/fiberseq\/library-prep\/\">Continue reading <span class=\"screen-reader-text\">Library Prep<\/span><\/a><\/div>\n","protected":false},"author":2,"featured_media":0,"parent":320,"menu_order":4,"comment_status":"closed","ping_status":"closed","template":"","meta":{"_acf_changed":false,"footnotes":""},"class_list":["post-381","page","type-page","status-publish","hentry"],"acf":[],"_links":{"self":[{"href":"https:\/\/nwgc.gs.washington.edu\/wordpress\/wp-json\/wp\/v2\/pages\/381","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/nwgc.gs.washington.edu\/wordpress\/wp-json\/wp\/v2\/pages"}],"about":[{"href":"https:\/\/nwgc.gs.washington.edu\/wordpress\/wp-json\/wp\/v2\/types\/page"}],"author":[{"embeddable":true,"href":"https:\/\/nwgc.gs.washington.edu\/wordpress\/wp-json\/wp\/v2\/users\/2"}],"replies":[{"embeddable":true,"href":"https:\/\/nwgc.gs.washington.edu\/wordpress\/wp-json\/wp\/v2\/comments?post=381"}],"version-history":[{"count":0,"href":"https:\/\/nwgc.gs.washington.edu\/wordpress\/wp-json\/wp\/v2\/pages\/381\/revisions"}],"up":[{"embeddable":true,"href":"https:\/\/nwgc.gs.washington.edu\/wordpress\/wp-json\/wp\/v2\/pages\/320"}],"wp:attachment":[{"href":"https:\/\/nwgc.gs.washington.edu\/wordpress\/wp-json\/wp\/v2\/media?parent=381"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}