{"id":1503,"date":"2021-03-07T12:29:30","date_gmt":"2021-03-07T11:29:30","guid":{"rendered":"http:\/\/directquant.eu\/?p=1503"},"modified":"2021-03-07T13:14:00","modified_gmt":"2021-03-07T12:14:00","slug":"dna-and-rna-extraction-from-swabs","status":"publish","type":"post","link":"https:\/\/directquant.eu\/?p=1503","title":{"rendered":"DNA and RNA extraction from swabs"},"content":{"rendered":"<p>This protocol is designed and validated for quantitative extraction of bacteria, viruses (DNA and RNA) and epithelial cells collected with swabs in less than 5 minutes using a single tube. If the aim of the study is bacterial or virus DNA\/RNA detection\/quantification genomic DNA from epithelial cells within the sample can be used as positive control for correct sample collection and extraction.<\/p>\n\n\n<p> <br> <\/p>\n\n\n<p>Additional equipment:<\/p>\n<ul>\n<li style=\"margin-bottom: 1rem\">Snap tool or scissors<\/li>\n<li style=\"margin-bottom: 1rem\">Thermoblock (with agitation is preferred) or cup of boiling water<\/li>\n<li style=\"margin-bottom: 1rem\">Centrifuge reaching 10.000g. (If the centrifuge is reaching only 3.000g the centrifugation time should be increased as indicated below).<\/li>\n<\/ul>\n<p>The protocol is identical for <a href=\"https:\/\/directquant.eu\/?page_id=340\"><strong>DireCtQuant 100ST (#DCQ100ST)<\/strong><\/a> and <a href=\"https:\/\/directquant.eu\/?page_id=344\"><strong>DireCtQuant 100W (#DCQ100W)<\/strong><\/a>.<\/p>\n\n\n<p> <br> <\/p>\n\n\n<p>For illustration purposes <a href=\"https:\/\/directquant.eu\/?page_id=340\"><strong>#DCQ100ST<\/strong><\/a> is used.<\/p>\n\n\n<p> <br> <\/p>\n\n\n\n<figure class=\"wp-block-image size-large\"><img loading=\"lazy\" width=\"593\" height=\"467\" src=\"https:\/\/directquant.eu\/wp-content\/uploads\/2021\/03\/swab_extraction.png\" alt=\"\" class=\"wp-image-1504\" srcset=\"https:\/\/directquant.eu\/wp-content\/uploads\/2021\/03\/swab_extraction.png 593w, https:\/\/directquant.eu\/wp-content\/uploads\/2021\/03\/swab_extraction-300x236.png 300w\" sizes=\"(max-width: 593px) 100vw, 593px\" \/><\/figure>\n\n\n<ol type=\"1\">\n<li style=\"margin-bottom: 1rem\">Pipet 200 \u03bcl of <strong><a rel=\"noreferrer noopener\" href=\"https:\/\/directquant.eu\/?page_id=340\" target=\"_blank\">#DCQ100ST<\/a><\/strong> or <strong><a rel=\"noreferrer noopener\" href=\"https:\/\/directquant.eu\/?page_id=344\" target=\"_blank\">#DCQ100W<\/a><\/strong> in a 1.5 ml tube.<\/li>\n<li style=\"margin-bottom: 1rem\">Snip the swab applicator as high as possible, permitting the tube to be closed.<\/li>\n<li style=\"margin-bottom: 1rem\">Close the tube. Make sure that the swab is completely wetted by the reagent.<\/li>\n<li style=\"margin-bottom: 1rem\">Incubate 3 minutes at 90\u00b0C. If possible, apply agitation during the incubation.<\/li>\n<li style=\"margin-bottom: 1rem\">Leave the tube 1 minute on the bench to cool down.<\/li>\n<li style=\"margin-bottom: 1rem\">With a sterile tool reverse the swab upside down.<\/li>\n<li style=\"margin-bottom: 1rem\">Close the tube and centrifuge at 10.000g for 30s in order to collect all the liquid absorbed in the swab. For Minicentrifuge reaching only 3.000g increase time to 3 minutes.<\/li>\n<li style=\"margin-bottom: 1rem\">Discard the swab.<\/li>\n<li style=\"margin-bottom: 1rem\">The samples can be stored and transported at room temperature after the extraction. For more information about the lon-term storage conditions check <strong><a rel=\"noreferrer noopener\" href=\"https:\/\/directquant.eu\/?page_id=340\" target=\"_blank\">Dire<em>Ct<\/em>Quant 100ST (#DCQ100ST)<\/a><\/strong> or <strong><a rel=\"noreferrer noopener\" href=\"https:\/\/directquant.eu\/?page_id=344\" target=\"_blank\">Dire<em>Ct<\/em>Quant 100W (#DCQ100W)<\/a><\/strong> protocols respectively.<\/li>\n<li style=\"margin-bottom: 1rem\">Use 1.0 \u03bcl of the sample per 20.0 \u03bcl PCR reaction. The extracted sample should be enough for around 200 detections. For more details about the PCR reaction conditions check <strong><a rel=\"noreferrer noopener\" href=\"https:\/\/directquant.eu\/?page_id=340\" target=\"_blank\">Dire<em>Ct<\/em>Quant 100ST (#DCQ100ST)<\/a><\/strong> or <strong><a rel=\"noreferrer noopener\" href=\"https:\/\/directquant.eu\/?page_id=344\" target=\"_blank\">Dire<em>Ct<\/em>Quant 100W (#DCQ100W)<\/a><\/strong> protocols respectively.<\/li>\n<\/ol>\n<p><!-- \/wp:list --><\/p>\n<p><!-- wp:separator --><\/p>\n<hr class=\"wp-block-separator\">\n<!-- \/wp:separator --><p><\/p>\n<p><!-- wp:paragraph --><\/p>\n<p><a href=\"https:\/\/directquant.eu\/wp-content\/uploads\/2021\/03\/DCQ100ST_DCQ100W_Swab_extraction.pdf\" target=\"_blank\" rel=\"noopener noreferrer\">Download as PDF file<\/a><\/p>\n<p><!-- \/wp:paragraph --><\/p>","protected":false},"excerpt":{"rendered":"<p>This protocol is designed and validated for quantitative extraction of bacteria, viruses (DNA and RNA) and epithelial cells collected with swabs in less than 5 minutes using a single tube. If the aim of the study is bacterial or virus DNA\/RNA detection\/quantification genomic DNA from epithelial cells within the sample can be used as positive [&hellip;]<\/p>\n","protected":false},"author":2,"featured_media":1504,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"jetpack_publicize_message":"","jetpack_is_tweetstorm":false,"jetpack_publicize_feature_enabled":true},"categories":[12,10],"tags":[22,20,24,21,18,23],"jetpack_publicize_connections":[],"jetpack_featured_media_url":"https:\/\/directquant.eu\/wp-content\/uploads\/2021\/03\/swab_extraction.png","jetpack_sharing_enabled":true,"jetpack_shortlink":"https:\/\/wp.me\/p91V39-of","_links":{"self":[{"href":"https:\/\/directquant.eu\/index.php?rest_route=\/wp\/v2\/posts\/1503"}],"collection":[{"href":"https:\/\/directquant.eu\/index.php?rest_route=\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/directquant.eu\/index.php?rest_route=\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/directquant.eu\/index.php?rest_route=\/wp\/v2\/users\/2"}],"replies":[{"embeddable":true,"href":"https:\/\/directquant.eu\/index.php?rest_route=%2Fwp%2Fv2%2Fcomments&post=1503"}],"version-history":[{"count":3,"href":"https:\/\/directquant.eu\/index.php?rest_route=\/wp\/v2\/posts\/1503\/revisions"}],"predecessor-version":[{"id":1510,"href":"https:\/\/directquant.eu\/index.php?rest_route=\/wp\/v2\/posts\/1503\/revisions\/1510"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/directquant.eu\/index.php?rest_route=\/wp\/v2\/media\/1504"}],"wp:attachment":[{"href":"https:\/\/directquant.eu\/index.php?rest_route=%2Fwp%2Fv2%2Fmedia&parent=1503"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/directquant.eu\/index.php?rest_route=%2Fwp%2Fv2%2Fcategories&post=1503"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/directquant.eu\/index.php?rest_route=%2Fwp%2Fv2%2Ftags&post=1503"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}