{"id":1428,"date":"2023-02-14T01:52:55","date_gmt":"2023-02-13T16:52:55","guid":{"rendered":"https:\/\/bdsl.jbnu.ac.kr\/blog\/?p=1428"},"modified":"2023-11-21T15:58:21","modified_gmt":"2023-11-21T06:58:21","slug":"amplicon-sequencing","status":"publish","type":"post","link":"https:\/\/bdsl.jbnu.ac.kr\/blog\/amplicon-sequencing\/","title":{"rendered":"Amplicon sequencing"},"content":{"rendered":"\t\t\t\t<div class=\"wp-block-uagb-table-of-contents uagb-toc__align-left uagb-toc__columns-1  uagb-block-e4df075a      \"\n\t\t\t\t\tdata-scroll= \"1\"\n\t\t\t\t\tdata-offset= \"30\"\n\t\t\t\t\tstyle=\"\"\n\t\t\t\t>\n\t\t\t\t<div class=\"uagb-toc__wrap\">\n\t\t\t\t\t\t<div class=\"uagb-toc__title\">\n\t\t\t\t\t\t\tTable Of Contents\t\t\t\t\t\t<\/div>\n\t\t\t\t\t\t\t\t\t\t\t\t\t\t\t\t\t\t\t\t\t\t<div class=\"uagb-toc__list-wrap \">\n\t\t\t\t\t\t<ol class=\"uagb-toc__list\"><li class=\"uagb-toc__list\"><a href=\"#introduction\" class=\"uagb-toc-link__trigger\">Introduction<\/a><li class=\"uagb-toc__list\"><a href=\"#amplicon-primers-for-bacteria\" class=\"uagb-toc-link__trigger\">Amplicon primers for bacteria<\/a><ul class=\"uagb-toc__list\"><li class=\"uagb-toc__list\"><a href=\"#illumina-v3-v4\" class=\"uagb-toc-link__trigger\">Illumina (V3-V4)<\/a><\/li><\/ul><\/li><li class=\"uagb-toc__list\"><a href=\"#library-construction\" class=\"uagb-toc-link__trigger\">Library construction<\/a><li class=\"uagb-toc__list\"><a href=\"#amplicon-sequencing\" class=\"uagb-toc-link__trigger\">Amplicon sequencing<\/a><li class=\"uagb-toc__list\"><a href=\"#data-analysis\" class=\"uagb-toc-link__trigger\">Data analysis<\/a><ul class=\"uagb-toc__list\"><li class=\"uagb-toc__list\"><a href=\"#generate-microbial-feature-table\" class=\"uagb-toc-link__trigger\">Generate microbial feature table<\/a><li class=\"uagb-toc__list\"><li class=\"uagb-toc__list\"><a href=\"#taxonomy-annotation\" class=\"uagb-toc-link__trigger\">Taxonomy annotation<\/a><\/ul><\/ul><\/ol>\t\t\t\t\t<\/div>\n\t\t\t\t\t\t\t\t\t<\/div>\n\t\t\t\t<\/div>\n\t\t\t\n\n\n<h2 class=\"wp-block-heading\">Introduction<\/h2>\n\n\n\n<p class=\"wp-block-paragraph\"><strong>(\uae30\ubcf8\uac1c\ub150)<\/strong> Amplicon sequencing\uc740 PCR\uc744 \uc774\uc6a9\ud574 \uc99d\ud3ed\ud55c DNA\uc778 amplicon\uc758 \uc11c\uc5f4\uc744 \uacb0\uc815\ud558\ub294 \uc2e4\ud5d8\uc774\ub2e4. \uc774 \ubc29\ubc95\uc740 \ub2e4\uc591\ud55c \ubbf8\uc0dd\ubb3c\uc774 \uc11e\uc5ec \uc788\ub294 \ubbf8\uc0dd\ubb3c \uade0\ucd1d\uc5d0 \uc5b4\ub5a4 \ubbf8\uc0dd\ubb3c\uc774 \uc874\uc7ac\ud558\ub294\uc9c0, \uadf8\ub9ac\uace0 \uadf8 \uc0c1\ub300\uc801\uc778 \uc591\uc740 \uc5bc\ub9c8\uc778\uc9c0 \uce21\uc815\ud558\uae30 \uc704\ud574 \uc8fc\ub85c \uc774\uc6a9\ub41c\ub2e4. \uc138\uade0\uc740 \uadf8 \uc885\ub958\uc5d0 \ub530\ub77c \uc11c\ub85c \ub2e4\ub978 16S rRNA \uc720\uc804\uc790 \uc11c\uc5f4\uc744 \uc9c0\ub2c8\uace0 \uc788\uc73c\uba70, \ub3d9\uc2dc\uc5d0 16S rRNA\uc758 \uc77c\ubd80\ubd84\uc740 \ub9e4\uc6b0 \uc798 \ubcf4\uc874\ub41c\ub2e4\ub294 \ud2b9\uc131\uc744 \ud65c\uc6a9\ud55c \ubc29\ubc95\uc774\ub2e4. \ub9e4\uc6b0 \uc798 \ubcf4\uc874\ub418\ub294 \ub450 \ubd80\ubd84\uc5d0 \uacb0\ud569\ud558\ub294 primer\ub97c \uc774\uc6a9\ud574 PCR \ubc18\uc751\uc744 \uc9c4\ud589\ud558\uba74, \uadf8 \uc0ac\uc774\uc5d0 \uc788\ub294 \ubcc0\uc774\uac00 \ub192\uc740 \uc11c\uc5f4\uc744 \uc77d\uc744 \uc218 \uc788\uace0, \ud574\ub2f9 \uc11c\uc5f4\ub85c\ubd80\ud130 \ubbf8\uc0dd\ubb3c\uc758 \uc885\ub958\ub97c \ud2b9\uc815\ud560 \uc218 \uc788\ub2e4. \ub354 \ub098\uc544\uac00, \uac01\uac01\uc758 \uc11c\uc5f4\uc774 \uba87 \ubc88 \uad00\ucc30\ub418\ub294\uc9c0 \ubd84\uc11d\ud568\uc73c\ub85c\uc368 \ubd84\uc11d\ud55c \uc0d8\ud50c \uc548\uc5d0 \uc874\uc7ac\ud558\ub294 \ubbf8\uc0dd\ubb3c\uc758 \uc0c1\ub300\uc801\uc778 \uc591\uc744 \ubd84\uc11d\ud560 \uc218 \uc788\ub2e4.<\/p>\n\n\n\n<p class=\"wp-block-paragraph\"><strong>(\uac00\ubcc0\uc601\uc5ed)<\/strong> Illumina MiSeq \uacfc \uac19\uc740  NGS \ubc29\ubc95\uc744 \ud1b5\ud574 \uc77d\uc744 \uc218 \uc788\ub294 \uc11c\uc5f4\uc758 \uae38\uc774\uac00 500\uac1c \ubbf8\ub9cc\uc73c\ub85c \uc81c\ud55c\ub41c\ub2e4. \ub530\ub77c\uc11c bacterial 16S rRNA gene \uc11c\uc5f4 \uc804\uccb4\uac00 \uc544\ub2cc 16S rRNA \uc11c\uc5f4\uc758 variable region\uc744 PCR\uc744 \uc774\uc6a9\ud574 \uc99d\ud3ed\ud55c\ub2e4. Bacterial 16S rRNA\ub294 \uc798 \ubcf4\uc874\ub41c \uc9c0\uc5ed\uacfc \uc18d \ud639\uc740 \uc885 \ubcc4\ub85c \ucc28\uc774\ub97c \ubcf4\uc774\ub294 9\uac1c\uc758 \uac00\ubcc0 \uc9c0\uc5ed (variable region)\uc744 \uac00\uc9c0\uace0 \uc788\ub2e4. PCR\uc744 \uc774\uc6a9\ud574 variable region\uc758 \uc11c\uc5f4\uc744 \uc99d\ud3ed\ud568\uc73c\ub85c\uc368, \uc5b4\ub5a4 \ud658\uacbd \ud639\uc740 \uc0d8\ud50c \uc548\uc5d0 \uc874\uc7ac\ud558\ub294 \ubbf8\uc0dd\ubb3c\uc758 \uc870\uc131\uc744 \ubd84\uc11d\ud560 \uc218 \uc788\ub2e4.<\/p>\n\n\n\n<p class=\"wp-block-paragraph\">\uac00\ubcc0\uc601\uc5ed\uc758 \ubcc0\ud654\ub294 \ubbf8\uc0dd\ubb3c\uc758 \uc885\ub958\uc5d0 \ub530\ub77c \ucc28\uc774\ub97c \ubcf4\uc778\ub2e4. \ub530\ub77c\uc11c \uc99d\ud3ed\ud560 \uac00\ubcc0 \uc601\uc5ed \uc120\ud0dd\uc5d0 \ub530\ub77c \ubd84\ub9ac\ud560 \uc218 \uc788\ub294 \ubbf8\uc0dd\ubb3c\uc758 \uc885\ub958\uac00 \ub2ec\ub77c\uc9c4\ub2e4. \uc790\uc138\ud55c \uc0ac\ud56d\uc740 \ub2e4\uc74c \ub17c\ubb38\uc744 \ucc38\uace0\ud55c\ub2e4.<\/p>\n\n\n\n<div class=\"wp-block-uagb-container uagb-block-65c72198 alignfull uagb-is-root-container\"><div class=\"uagb-container-inner-blocks-wrap\">\n<p class=\"wp-block-paragraph\">\ucc38\uace0 \ub17c\ubb38: Johnson, J. S., et al. (2019). &#8220;Evaluation of 16S rRNA gene sequencing for species and strain-level microbiome analysis.&#8221; Nat Commun 10(1): 5029.<\/p>\n\n\n\n<div class=\"wp-block-file\"><a id=\"wp-block-file--media-132a65dd-8751-4fb3-bf47-2dc7ab2d0caa\" href=\"https:\/\/bdsl.jbnu.ac.kr\/blog\/wp-content\/uploads\/2023\/07\/s41467-019-13036-1.pdf\">s41467-019-13036-1<\/a><a href=\"https:\/\/bdsl.jbnu.ac.kr\/blog\/wp-content\/uploads\/2023\/07\/s41467-019-13036-1.pdf\" class=\"wp-block-file__button wp-element-button\" download aria-describedby=\"wp-block-file--media-132a65dd-8751-4fb3-bf47-2dc7ab2d0caa\">Download<\/a><\/div>\n<\/div><\/div>\n\n\n\n<h2 class=\"wp-block-heading\">Amplicon primers for bacteria<\/h2>\n\n\n\n<p class=\"wp-block-paragraph\">Klindworth et al.\ub294 <em>in silico<\/em> amplicon sequencing \uc744 \ud1b5\ud574 V3-V4 region\uc744 \ub300\uc0c1\uc73c\ub85c\ud55c \uc544\ub798\uc758 primer \uc11c\uc5f4\uc774 bacteria \ubd84\ub958\ub97c \uc704\ud574 \uac00\uc7a5 \uc88b\uc740 \uc11c\uc5f4\uc784\uc744 \ubc1c\uacac\ud558\uc600\ub2e4 (A.&nbsp;Klindworth, Nucleic Acids Res.,&nbsp;41&nbsp;(2012), p.&nbsp;e1). Primer\uc758 \uc11c\uc5f4\uc740 \uc544\ub798\uc640 \uac19\ub2e4. \uc5ec\ub7ec \uc5f0\uad6c\uc5d0\uc11c \uc774 \uc601\uc5ed\uc744 \ubd84\uc11d \ub300\uc0c1\uc73c\ub85c \uc0bc\uace0 \uc788\ub294 \uac83\uc744 \uc54c \uc218 \uc788\ub2e4. \ud558\uc9c0\ub9cc, \uac01 \uc5f0\uad6c\uc758 \ubaa9\uc801\uc5d0 \ub530\ub77c \ub2e4\ub978 \uac00\ubcc0 \ubd80\uc704\ub97c \uc120\ud0dd\ud558\ub294 \uacbd\uc6b0\uac00 \ub9ce\uc774 \uc788\uc73c\ubbc0\ub85c, \ub370\uc774\ud130 \ubd84\uc11d \uc2dc \uc5b4\ub5a4 \uc601\uc5ed\uc774 \ubd84\uc11d\uc5d0 \uc774\uc6a9\ub418\uc5c8\ub294\uc9c0 \ud655\uc778\ud560 \ud544\uc694\uac00 \uc788\ub2e4.<\/p>\n\n\n\n<figure class=\"wp-block-table\"><table><thead><tr><th>Region<\/th><th>Direction<\/th><th>Primer sequence<\/th><\/tr><\/thead><tbody><tr><td>v3-v4<\/td><td>Forward<\/td><td>CCTACGGGNGGCWGCAG<\/td><\/tr><tr><td>v3-v4<\/td><td>Reverse<\/td><td>GACTACHVGGGTATCTAATCC<\/td><\/tr><\/tbody><\/table><\/figure>\n\n\n\n<h3 class=\"wp-block-heading\">Illumina (V3-V4)<\/h3>\n\n\n\n<p class=\"wp-block-paragraph\">Klindworth\uac00 \uc81c\uc2dc\ud55c \uc11c\uc5f4\uc740 Illumina\uc5d0\uc11c\ub3c4 16S rRNA sequencing\uc744 \uc704\ud574 \ucd94\ucc9c\ud558\ub294 primer \uc870\ud569\uc774\ub2e4. Illumina\uc5d0\uc11c\ub294 Illumina sequencing\uc744 \uc704\ud55c overhang adapter\ub97c \ud3ec\ud568\ud55c primer\ub97c \ucd94\ucc9c\ud558\uace0 \uc788\uc73c\uba70, \ud544\uc694\uc5d0 \ub530\ub77c\uc11c\ub294 \uc784\uc758\uc758 primer \uc11c\uc5f4\uc744 \uc774\uc6a9\ud560 \uc218\ub3c4 \uc788\ub2e4. <\/p>\n\n\n\n<p class=\"wp-block-paragraph\"><a href=\"https:\/\/support.illumina.com\/documents\/documentation\/chemistry_documentation\/16s\/16s-metagenomic-library-prep-guide-15044223-b.pdf\">https:\/\/support.illumina.com\/documents\/documentation\/chemistry_documentation\/16s\/16s-metagenomic-library-prep-guide-15044223-b.pdf<\/a><\/p>\n\n\n\n<p class=\"wp-block-paragraph\"><strong>Primer sequence<\/strong><\/p>\n\n\n\n<ul class=\"wp-block-list\">\n<li>Forward: CCTACGGGNGGCWGCAG<\/li>\n\n\n\n<li>Reverse: GACTACHVGGGTATCTAATCC<\/li>\n<\/ul>\n\n\n\n<p class=\"wp-block-paragraph\"><strong>Primer sequence with overhang adapter<\/strong><\/p>\n\n\n\n<ul class=\"wp-block-list\">\n<li>Forward: TCGTCGGCAGCGTCAGATGTGTATAAGAGACAGCCTACGGGNGGCWGCAG<\/li>\n\n\n\n<li>Reverse: GTCTCGTGGGCTCGGAGATGTGTATAAGAGACAGGACTACHVGGGTATCTAATCC<\/li>\n<\/ul>\n\n\n\n<p class=\"wp-block-paragraph\"><strong>Overhang adapter<\/strong><\/p>\n\n\n\n<ul class=\"wp-block-list\">\n<li>Forward: TCGTCGGCAGCGTCAGATGTGTATAAGAGACAG\u2010[locus\u2010specificsequence]<\/li>\n\n\n\n<li>Reverse: GTCTCGTGGGCTCGGAGATGTGTATAAGAGACAG\u2010[locus\u2010specificsequence]<\/li>\n<\/ul>\n\n\n\n<h2 class=\"wp-block-heading\">Library construction<\/h2>\n\n\n\n<p class=\"wp-block-paragraph\">PCR \uacfc\uc815\uc744 \uac70\ucc98 \uc99d\ubcf5\ud55c DNA\ub97c library\ub77c \ud55c\ub2e4. \uc0d8\ud50c\uc5d0\uc11c genomic DNA\ub97c \ucd94\ucd9c\ud558\uace0, PCR \ubc18\uc751\uc744 \uc218\ud589\ud558\uace0, \uc99d\ud3ed\ud55c DNA\ub97c \ucd94\ucd9c\ud558\ub294 \uacfc\uc815\uc744 \uac70\uce5c\ub2e4. \uc2e4\ud5d8 \ubaa9\uc801\uc5d0 \ub530\ub77c DNA\ub97c \uc900\ube44\ud558\ub294 \uacfc\uc815\uc5d0\uc11c \ub9ce\uc740 \ucc28\uc774\uac00 \ubc1c\uc0dd\ud55c\ub2e4. \ub2e4\uc74c\uc740 \ud55c\uad6d\uc2dd\ud488\uc5f0\uad6c\uc6d0\uc5d0\uc11c \uc0ac\uc6a9\ud558\ub294 \ud504\ub85c\ud1a0\ucf5c\uc758 \uc77c\ubd80\uc774\ub2e4.<\/p>\n\n\n\n<ul class=\"wp-block-list\">\n<li>QIAamp DNA stool minikit<\/li>\n\n\n\n<li>Bead bitting and Lysis: Zirconia beads (BioSpec), ASL lysis buffer (Qiagen), 95 degrees 15 mins, bead beating 30Hz 1 min (Qiagen TissueLyser II)<\/li>\n\n\n\n<li>Extraction: QIAcub system (Qiagen)<\/li>\n<\/ul>\n\n\n\n<p class=\"wp-block-paragraph\">\uc2e4\ud5d8 \ubc29\ubc95\uc5d0 \ub300\ud574\uc11c\ub294 \ub2e4\uc74c \ub17c\ubb38\uc744 \ucc38\uace0\ud55c\ub2e4. <\/p>\n\n\n\n<div class=\"wp-block-uagb-container uagb-block-d87a6550 alignfull uagb-is-root-container\"><div class=\"uagb-container-inner-blocks-wrap\">\n<p class=\"wp-block-paragraph\">Lim, M. Y., et al. (2021). &#8220;Gut Microbiome Structure and Association with Host Factors in a Korean Population.&#8221; Msystems 6(4). <\/p>\n\n\n\n<div class=\"wp-block-file\"><a id=\"wp-block-file--media-8fb64081-7c5f-4156-bdc4-2844780137c8\" href=\"https:\/\/bdsl.jbnu.ac.kr\/blog\/wp-content\/uploads\/2023\/02\/Lim_et_al___2021___Gut_Microbiome_Structure_an.pdf\">Lim_et_al___2021___Gut_Microbiome_Structure_an<\/a><a href=\"https:\/\/bdsl.jbnu.ac.kr\/blog\/wp-content\/uploads\/2023\/02\/Lim_et_al___2021___Gut_Microbiome_Structure_an.pdf\" class=\"wp-block-file__button wp-element-button\" download aria-describedby=\"wp-block-file--media-8fb64081-7c5f-4156-bdc4-2844780137c8\">Download<\/a><\/div>\n<\/div><\/div>\n\n\n\n<h2 class=\"wp-block-heading\">Amplicon sequencing<\/h2>\n\n\n\n<h2 class=\"wp-block-heading\">Data analysis<\/h2>\n\n\n\n<p class=\"wp-block-paragraph\">Sequencing machine\uc5d0\uc11c \uc5bb\uc740 \uacb0\uacfc\ub97c \uac01 sample\uc5d0 \ub300\ud55c barcode\ub97c \uc774\uc6a9\ud558\uc5ec \ubd84\ub9ac(demultiplexing)\ud558\uba74, \uac01 \uc0d8\ud50c\uc5d0 \ub300\ud574 forward\uc640 reverse\uc758 \ub450 \uac1c\uc758 FASTQ format \ud30c\uc77c\uc744 \uc5bb\uc744 \uc218 \uc788\ub2e4 (paired-end sequencing).  <\/p>\n\n\n\n<h3 class=\"wp-block-heading\">Generate microbial feature table<\/h3>\n\n\n\n<figure class=\"wp-block-embed is-type-wp-embed is-provider-biomedical-data-science-laboratory wp-block-embed-biomedical-data-science-laboratory\"><div class=\"wp-block-embed__wrapper\">\n<blockquote class=\"wp-embedded-content\" data-secret=\"qg7ht4bo13\"><a href=\"https:\/\/bdsl.jbnu.ac.kr\/blog\/generate-microbial-feature-table\/\">Generate microbial feature table<\/a><\/blockquote><iframe loading=\"lazy\" class=\"wp-embedded-content\" sandbox=\"allow-scripts\" security=\"restricted\" style=\"position: absolute; clip: rect(1px, 1px, 1px, 1px);\" title=\"&#8220;Generate microbial feature table&#8221; &#8212; Biomedical Data Science Laboratory\" src=\"https:\/\/bdsl.jbnu.ac.kr\/blog\/generate-microbial-feature-table\/embed\/#?secret=8rPHvgiJvl#?secret=qg7ht4bo13\" data-secret=\"qg7ht4bo13\" width=\"500\" height=\"282\" frameborder=\"0\" marginwidth=\"0\" marginheight=\"0\" scrolling=\"no\"><\/iframe>\n<\/div><\/figure>\n\n\n\n<h3 class=\"wp-block-heading\">Taxonomy annotation<\/h3>\n\n\n\n<figure class=\"wp-block-embed is-type-wp-embed is-provider-biomedical-data-science-laboratory wp-block-embed-biomedical-data-science-laboratory\"><div class=\"wp-block-embed__wrapper\">\n<blockquote class=\"wp-embedded-content\" data-secret=\"ks2pzB9C71\"><a href=\"https:\/\/bdsl.jbnu.ac.kr\/blog\/taxonomy-annotation-for-amplicon-sequencing\/\">Taxonomy annotation for amplicon sequencing<\/a><\/blockquote><iframe loading=\"lazy\" class=\"wp-embedded-content\" sandbox=\"allow-scripts\" security=\"restricted\" style=\"position: absolute; clip: rect(1px, 1px, 1px, 1px);\" title=\"&#8220;Taxonomy annotation for amplicon sequencing&#8221; &#8212; Biomedical Data Science Laboratory\" src=\"https:\/\/bdsl.jbnu.ac.kr\/blog\/taxonomy-annotation-for-amplicon-sequencing\/embed\/#?secret=lTp1RGCzIY#?secret=ks2pzB9C71\" data-secret=\"ks2pzB9C71\" width=\"500\" height=\"282\" frameborder=\"0\" marginwidth=\"0\" marginheight=\"0\" scrolling=\"no\"><\/iframe>\n<\/div><\/figure>\n","protected":false},"excerpt":{"rendered":"<p>Introduction (\uae30\ubcf8\uac1c\ub150) Amplicon sequencing\uc740 PCR\uc744 \uc774\uc6a9\ud574 \uc99d\ud3ed\ud55c DNA\uc778 amplicon\uc758 \uc11c\uc5f4\uc744 \uacb0\uc815\ud558\ub294 \uc2e4\ud5d8\uc774\ub2e4. \uc774 \ubc29\ubc95\uc740 \ub2e4\uc591\ud55c \ubbf8\uc0dd\ubb3c\uc774 \uc11e\uc5ec \uc788\ub294 \ubbf8\uc0dd\ubb3c \uade0\ucd1d\uc5d0 \uc5b4\ub5a4 \ubbf8\uc0dd\ubb3c\uc774 \uc874\uc7ac\ud558\ub294\uc9c0, \uadf8\ub9ac\uace0 \uadf8 \uc0c1\ub300\uc801\uc778 \uc591\uc740 \uc5bc\ub9c8\uc778\uc9c0 \uce21\uc815\ud558\uae30 \uc704\ud574 \uc8fc\ub85c \uc774\uc6a9\ub41c\ub2e4. \uc138\uade0\uc740 \uadf8 \uc885\ub958\uc5d0 \ub530\ub77c \uc11c\ub85c \ub2e4\ub978 16S rRNA \uc720\uc804\uc790 \uc11c\uc5f4\uc744 \uc9c0\ub2c8\uace0 \uc788\uc73c\uba70, \ub3d9\uc2dc\uc5d0 16S rRNA\uc758 \uc77c\ubd80\ubd84\uc740 \ub9e4\uc6b0 \uc798 \ubcf4\uc874\ub41c\ub2e4\ub294 \ud2b9\uc131\uc744 \ud65c\uc6a9\ud55c \ubc29\ubc95\uc774\ub2e4. \ub9e4\uc6b0 [&hellip;]<\/p>\n","protected":false},"author":1,"featured_media":0,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"_uag_custom_page_level_css":"","site-sidebar-layout":"default","site-content-layout":"","ast-site-content-layout":"default","site-content-style":"default","site-sidebar-style":"default","ast-global-header-display":"","ast-banner-title-visibility":"","ast-main-header-display":"","ast-hfb-above-header-display":"","ast-hfb-below-header-display":"","ast-hfb-mobile-header-display":"","site-post-title":"","ast-breadcrumbs-content":"","ast-featured-img":"","footer-sml-layout":"","ast-disable-related-posts":"","theme-transparent-header-meta":"","adv-header-id-meta":"","stick-header-meta":"","header-above-stick-meta":"","header-main-stick-meta":"","header-below-stick-meta":"","astra-migrate-meta-layouts":"set","ast-page-background-enabled":"default","ast-page-background-meta":{"desktop":{"background-color":"var(--ast-global-color-4)","background-image":"","background-repeat":"repeat","background-position":"center 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