{"id":152,"date":"2016-09-26T12:24:46","date_gmt":"2016-09-26T04:24:46","guid":{"rendered":"http:\/\/140.109.28.108\/brcasngscore\/?page_id=152"},"modified":"2016-10-03T15:47:57","modified_gmt":"2016-10-03T07:47:57","slug":"sample-preparation","status":"publish","type":"page","link":"https:\/\/ngs.biodiv.tw\/NGSCore\/sample-preparation\/","title":{"rendered":"Sample Preparation"},"content":{"rendered":"<h1><span style=\"color: #7aa200;\">Preparation of Nucleic Acid<\/span><\/h1>\n<p><span style=\"font-size: 14pt; color: #404040;\">Nucleic acid extraction by either organic method or commercial kit requires at least twice ethanol wash steps. Do additional spin to collect residual solution at tube bottom and pipet to remove it. Air dry on benchtop (preferred over speedvac). Chemicals and contaminants (e.g. polysaccharides, lipids, or pigment) should be removed to avoid inhibition on library construction.<\/span><\/p>\n<p><span style=\"font-size: 14pt; color: #404040;\">We recommend including a DNase step (in the RNA isolation) or a RNase step (in the DNA isolation), followed by phenol:chloroform purification, to increase the sample purity. Degraded RNA may be present after RNase treatment and visualized by 2% gel check. Some commercial kits provide on-column digest to help wash away degraded RNA.<\/span><\/p>\n<p><span style=\"font-size: 14pt; color: #404040;\">RNase inhibitor is generally NOT recommended. But if necessary, please add only at low concentration (e.g. half or quarter of the commercial suggestion) into the RNA sample, and indicate the brand name and the usage amount on the Sample Submission Form.<\/span><\/p>\n<p><span style=\"font-size: 14pt; color: #404040;\">Please dissolved the sample in <strong>nuclease-free molecular grade water<\/strong> or <strong>EB buffer (10 mM Tris-HCl, pH 8.0)<\/strong>, and make sure to be free of contamination.<\/span><\/p>\n<p><span style=\"font-size: 14pt; color: #404040;\">Please note <strong>an additional charge for sample purification<\/strong> will occur should there be concern of contamination (e.g. CTAB, polysugar, phenolic compound, etc).<\/span><\/p>\n","protected":false},"excerpt":{"rendered":"<p>Preparation of Nucleic Acid Nucleic acid extraction by  [&hellip;]<\/p>\n","protected":false},"author":1,"featured_media":0,"parent":0,"menu_order":0,"comment_status":"closed","ping_status":"closed","template":"","meta":{"footnotes":""},"class_list":["post-152","page","type-page","status-publish","hentry"],"_links":{"self":[{"href":"https:\/\/ngs.biodiv.tw\/NGSCore\/wp-json\/wp\/v2\/pages\/152","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/ngs.biodiv.tw\/NGSCore\/wp-json\/wp\/v2\/pages"}],"about":[{"href":"https:\/\/ngs.biodiv.tw\/NGSCore\/wp-json\/wp\/v2\/types\/page"}],"author":[{"embeddable":true,"href":"https:\/\/ngs.biodiv.tw\/NGSCore\/wp-json\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"https:\/\/ngs.biodiv.tw\/NGSCore\/wp-json\/wp\/v2\/comments?post=152"}],"version-history":[{"count":6,"href":"https:\/\/ngs.biodiv.tw\/NGSCore\/wp-json\/wp\/v2\/pages\/152\/revisions"}],"predecessor-version":[{"id":394,"href":"https:\/\/ngs.biodiv.tw\/NGSCore\/wp-json\/wp\/v2\/pages\/152\/revisions\/394"}],"wp:attachment":[{"href":"https:\/\/ngs.biodiv.tw\/NGSCore\/wp-json\/wp\/v2\/media?parent=152"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}