{"id":1323,"date":"2013-04-10T13:14:44","date_gmt":"2013-04-10T17:14:44","guid":{"rendered":"http:\/\/www.ambrosiasymbiosis.org\/labprotocols\/extraction-from-beetles\/"},"modified":"2015-10-19T08:46:11","modified_gmt":"2015-10-19T12:46:11","slug":"extraction-from-beetles","status":"publish","type":"post","link":"https:\/\/www.ambrosiasymbiosis.org\/labprotocols\/extraction-from-beetles\/","title":{"rendered":"Extraction from Beetles"},"content":{"rendered":"<h2>Protocol<\/h2>\n<h3>Materials<\/h3>\n<ul>\n<li>micropipette<\/li>\n<li>micropipette tips<\/li>\n<li>forceps<\/li>\n<li>scalpel<\/li>\n<li>razor<\/li>\n<li>petri dishes<\/li>\n<li>ethanol in petri dish<\/li>\n<\/ul>\n<h3>Procedure<\/h3>\n<p><b>Beetles:<\/b>\u00a010 female beetles<\/p>\n<ol>\n<li>Remove elytra and clip wings.<\/li>\n<li>Put each beetle in\u00a0<b>0.5 ml 1X PBS<\/b><\/li>\n<li>Vortex (or sonicate) and keep\/plate solution.<\/li>\n<li>In hood, separate beetle abdomen, head, and pronotum<\/li>\n<li>Glue abdomen onto a sterile plate, let dry in hood briefly.<\/li>\n<li>Meanwhile, put head in\u00a0<b>0.5 ml 1X PBS<\/b><\/li>\n<li>Crush (substitute for extracting mandibular pouches)<\/li>\n<li>Open up the abdomen, suck up the contents under metanotum, transfer it in\u00a0<i>0.5 ml 1X PBS&#8217;<\/i><\/li>\n<li>Remove gut, put in\u00a0<b>0.5 ml 1X PBS<\/b><\/li>\n<li>and crush.<\/li>\n<li>remove contents from mycangium and transfer in\u00a0<b>0.5 ml 1X PBS<\/b>.<\/li>\n<\/ol>\n<p><b>Sonicator test:<\/b>\u00a0use six beetles, sonicate two (separately) for 10, 30, 60 seconds. proceed as above. \u2013 NO DIFFERENCE, BEETLE STILL LIVING!<\/p>\n<p><b>Bead beater<\/b>\u00a0\u2013 fill tubes with 500 \u03bcl of PBS. More liquid prevents movement of beads.<\/p>\n","protected":false},"excerpt":{"rendered":"<p>Protocol Materials micropipette micropipette tips forceps scalpel razor petri dishes ethanol in petri dish Procedure Beetles:\u00a010 female beetles Remove elytra and clip wings. Put each beetle in\u00a00.5 ml 1X PBS Vortex (or sonicate) and keep\/plate solution. In hood, separate beetle abdomen, head, and pronotum Glue abdomen onto a sterile plate, let dry in hood briefly. [&hellip;]<\/p>\n","protected":false},"author":7,"featured_media":0,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"_s2mail":"yes","footnotes":""},"categories":[6],"tags":[],"class_list":["post-1323","post","type-post","status-publish","format-standard","hentry","category-beetles"],"_links":{"self":[{"href":"https:\/\/www.ambrosiasymbiosis.org\/labprotocols\/wp-json\/wp\/v2\/posts\/1323","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/www.ambrosiasymbiosis.org\/labprotocols\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/www.ambrosiasymbiosis.org\/labprotocols\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/www.ambrosiasymbiosis.org\/labprotocols\/wp-json\/wp\/v2\/users\/7"}],"replies":[{"embeddable":true,"href":"https:\/\/www.ambrosiasymbiosis.org\/labprotocols\/wp-json\/wp\/v2\/comments?post=1323"}],"version-history":[{"count":1,"href":"https:\/\/www.ambrosiasymbiosis.org\/labprotocols\/wp-json\/wp\/v2\/posts\/1323\/revisions"}],"predecessor-version":[{"id":1833,"href":"https:\/\/www.ambrosiasymbiosis.org\/labprotocols\/wp-json\/wp\/v2\/posts\/1323\/revisions\/1833"}],"wp:attachment":[{"href":"https:\/\/www.ambrosiasymbiosis.org\/labprotocols\/wp-json\/wp\/v2\/media?parent=1323"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/www.ambrosiasymbiosis.org\/labprotocols\/wp-json\/wp\/v2\/categories?post=1323"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/www.ambrosiasymbiosis.org\/labprotocols\/wp-json\/wp\/v2\/tags?post=1323"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}