{"id":597,"date":"2012-07-17T23:06:17","date_gmt":"2012-07-18T03:06:17","guid":{"rendered":"http:\/\/blogs.vassar.edu\/diffractionsymmetries\/?p=597"},"modified":"2013-06-14T10:26:36","modified_gmt":"2013-06-14T14:26:36","slug":"growing-c-elegans","status":"publish","type":"post","link":"https:\/\/pages.vassar.edu\/diffractionsymmetries\/2012\/07\/17\/growing-c-elegans\/","title":{"rendered":"Growing C.elegans"},"content":{"rendered":"<p>When growing <em>C. elegans, <\/em>large amounts can be grown in different mediums. The most popular is liquid medium. <em>C. elegans<\/em> are usually grown on S Medium using concentrated doses of \u00a0<em>E. coli<\/em> bacteria as their food. \u00a0Starting off, it is wise to grow just one generation of worms before harvesting them. After the worms produce multiple generations, harvesting should be done.<\/p>\n<p>One should make a concentrated plate of bacteria. This plate can be store around 4\/5\u00b0C for several weeks. One can store the plates in a subzero freezer (-70\u00b0C) indefinitely. Generally speaking, it is a sound idea to have ample amounts of \u00a0<em>E. coli<\/em> available for the C.elegans. However, the amount of food the worms will need will depend on the starting inoculate of worms and the length of time the worms are grown.<\/p>\n<p>Here are some of the common reagents used in growing C.elegans<\/p>\n<p><em>Reagents<\/em><\/p>\n<ul>\n<li>\u00a0NaCl, K<sub>2<\/sub> HPO<sub>4<\/sub>, 6 g KH<sub>2<\/sub>PO<sub>4<\/sub>, 1 ml cholesterol , water. Sterilize it\u00a0 by using an autoclave machine.<\/li>\n<li>1 M Potassium citrate pH 6.0<\/li>\n<li>Some trace metals solutions<\/li>\n<li>1 M CaCl<sub>2<\/sub><\/li>\n<li>2 or 3\u00a0 large plates of <em>C. elegans<\/em><\/li>\n<li>food, ie. concentrated\u00a0 <em>E. coli<\/em><\/li>\n<\/ul>\n<p>Procedures<\/p>\n<p>Inoculate the Medium with a does of\u00a0 <em>E. coli<\/em>\u00a0 from an previously made culture. Make sure you wash each plates of <em>C. elegans<\/em> properly. Shake the flask using a shaker at 20\u00b0C for a predetermined amount of time. Make sure the flask is shaken vigorously to ensure the culture is well oxygenated. Monitor each culture by examining them under a microscope. When examining the <em>C.elegans,<\/em>\u00a0if their food supply is depleted, it is time to replenish the supply. However one must also harvest them, which is usually done\u00a0 on the 5<sup>th<\/sup> day. Sterilize the harvesting tool, and transfer 5 or so <em>C.elegans<\/em> onto a new plate<\/p>\n","protected":false},"excerpt":{"rendered":"<p>When growing C. elegans, large amounts can be grown in different mediums. The most popular is liquid medium. C. elegans are usually grown on S Medium using concentrated doses of \u00a0E. coli bacteria as their food. \u00a0Starting off, it is &hellip; <a href=\"https:\/\/pages.vassar.edu\/diffractionsymmetries\/2012\/07\/17\/growing-c-elegans\/\">Continue reading <span class=\"meta-nav\">&rarr;<\/span><\/a><\/p>\n","protected":false},"author":2480,"featured_media":0,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[1],"tags":[],"class_list":["post-597","post","type-post","status-publish","format-standard","hentry","category-uncategorized"],"_links":{"self":[{"href":"https:\/\/pages.vassar.edu\/diffractionsymmetries\/wp-json\/wp\/v2\/posts\/597","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/pages.vassar.edu\/diffractionsymmetries\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/pages.vassar.edu\/diffractionsymmetries\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/pages.vassar.edu\/diffractionsymmetries\/wp-json\/wp\/v2\/users\/2480"}],"replies":[{"embeddable":true,"href":"https:\/\/pages.vassar.edu\/diffractionsymmetries\/wp-json\/wp\/v2\/comments?post=597"}],"version-history":[{"count":10,"href":"https:\/\/pages.vassar.edu\/diffractionsymmetries\/wp-json\/wp\/v2\/posts\/597\/revisions"}],"predecessor-version":[{"id":705,"href":"https:\/\/pages.vassar.edu\/diffractionsymmetries\/wp-json\/wp\/v2\/posts\/597\/revisions\/705"}],"wp:attachment":[{"href":"https:\/\/pages.vassar.edu\/diffractionsymmetries\/wp-json\/wp\/v2\/media?parent=597"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/pages.vassar.edu\/diffractionsymmetries\/wp-json\/wp\/v2\/categories?post=597"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/pages.vassar.edu\/diffractionsymmetries\/wp-json\/wp\/v2\/tags?post=597"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}