{"id":176,"date":"2016-08-18T16:40:52","date_gmt":"2016-08-18T16:40:52","guid":{"rendered":"http:\/\/baliga.systemsbiology.net\/see-interns\/hs2016\/?page_id=176"},"modified":"2016-10-13T00:29:28","modified_gmt":"2016-10-13T00:29:28","slug":"halobacterium-salinaru","status":"publish","type":"page","link":"https:\/\/baliga.systemsbiology.net\/see-interns\/hs2016\/projects\/cell-dynamics\/model-organism-halobacterium-salinarum-strain-nrc-1\/halobacterium-salinaru\/","title":{"rendered":"Finding Low Cost Substitutes for Ingredients used in Complete Media for Halobacterium salinarum"},"content":{"rendered":"<p style=\"text-align: justify\"><strong>By Sumaiya Sathar<\/strong><\/p>\n<h4 style=\"text-align: justify\">Introduction<\/h4>\n<p style=\"text-align: justify\"><b><\/b><b><\/b>Halobacterium salinarum is a type of archaea that resides in highly salty environments. H. salinarum is widely used as a model organism since their genetic processing method is very similar to that of humans. In a laboratory setting, halobacterium is grown in Complete Media which is a combination of laboratory-grade chemicals and proteins that provide the nutrients H. salinarum needs in order to survive.\u00a0<i>Halobacterium<\/i> is extremely simple to conduct experiments with and safely handle, thus making it a favorite amongst teachers around the world. The ingredients used in complete media are expensive and inaccessible for many teachers and aspiring scientists, so our goal is to find accessible and low cost ingredients to use as substitutes for the more expensive chemicals in complete media. Throughout the Internship my partners Sara, Nida and I have been testing various substitutes that teachers could use to prepare low cost media.<\/p>\n<h4 style=\"text-align: justify\">Materials and Methods<\/h4>\n<p style=\"text-align: justify\">To conduct these tests:<\/p>\n<p style=\"text-align: justify\">1.Substitute one ingredient from standard CM. In each case, we began by substituting the same mass of the substitute ingredient as is used in standard media.<\/p>\n<p style=\"text-align: justify\">2.Complete a growth curve experiment.<\/p>\n<p style=\"text-align: justify;padding-left: 30px\">a.Add 5 mL of the media to four separate 14 mL falcon tubes.<\/p>\n<p style=\"text-align: justify;padding-left: 30px\">b.Add 100 uL of wild type <i>Halobacterium<\/i> to three of the tubes. \u00a0Do not add cells to the fourth tube &#8211; this tube will serve as your control.<\/p>\n<p style=\"text-align: justify;padding-left: 30px\">c.Place the falcon tubes in a shaking incubator (220 rpm) at 37\u00b0C with light.<\/p>\n<p style=\"text-align: justify;padding-left: 30px\">d.Take the optical density (OD) of the sample every 4 hours for a 48 hour time period. When measuring OD, use absorbance at 600 nm.<\/p>\n<p style=\"text-align: justify;padding-left: 30px\">e.Graph the results and formulate our conclusions.<\/p>\n<p style=\"text-align: justify\">Below is a chart of all the ingredients found in regular CM media and the substitutes we tested.<\/p>\n<p style=\"text-align: justify\"><a href=\"https:\/\/baliga.systemsbiology.net\/see-interns\/hs2016\/wp-content\/uploads\/sites\/4\/2016\/08\/sumaiyasnip.png\"><img loading=\"lazy\" decoding=\"async\" class=\"alignnone wp-image-186\" src=\"https:\/\/baliga.systemsbiology.net\/see-interns\/hs2016\/wp-content\/uploads\/sites\/4\/2016\/08\/sumaiyasnip-300x152.png\" alt=\"sumaiyasnip\" width=\"491\" height=\"249\" srcset=\"https:\/\/baliga.systemsbiology.net\/see-interns\/hs2016\/wp-content\/uploads\/sites\/4\/2016\/08\/sumaiyasnip-300x152.png 300w, https:\/\/baliga.systemsbiology.net\/see-interns\/hs2016\/wp-content\/uploads\/sites\/4\/2016\/08\/sumaiyasnip.png 385w\" sizes=\"auto, (max-width: 491px) 100vw, 491px\" \/><\/a><\/p>\n<p><a href=\"https:\/\/baliga.systemsbiology.net\/see-interns\/hs2016\/wp-content\/uploads\/sites\/4\/2016\/08\/Halo-in-CM-growth-curve.png\"><img loading=\"lazy\" decoding=\"async\" class=\"alignnone wp-image-1008\" src=\"https:\/\/baliga.systemsbiology.net\/see-interns\/hs2016\/wp-content\/uploads\/sites\/4\/2016\/08\/Halo-in-CM-growth-curve-300x194.png\" alt=\"Halo in CM growth curve\" width=\"491\" height=\"317\" srcset=\"https:\/\/baliga.systemsbiology.net\/see-interns\/hs2016\/wp-content\/uploads\/sites\/4\/2016\/08\/Halo-in-CM-growth-curve-300x194.png 300w, https:\/\/baliga.systemsbiology.net\/see-interns\/hs2016\/wp-content\/uploads\/sites\/4\/2016\/08\/Halo-in-CM-growth-curve-768x497.png 768w, https:\/\/baliga.systemsbiology.net\/see-interns\/hs2016\/wp-content\/uploads\/sites\/4\/2016\/08\/Halo-in-CM-growth-curve.png 996w\" sizes=\"auto, (max-width: 491px) 100vw, 491px\" \/><\/a><\/p>\n<p style=\"text-align: justify\">Above is a graph of the growth curve for <i>Halobacterium<\/i> in regular complete media, without any substitutes.\u00a0The growth curves for Halobacterium in media with\u00a0substitutes were\u00a0compared to this growth curve in order to determine if the substitute did indeed work or not.<\/p>\n<h4 style=\"text-align: justify\">Results<\/h4>\n<p><a href=\"https:\/\/baliga.systemsbiology.net\/see-interns\/hs2016\/wp-content\/uploads\/sites\/4\/2016\/08\/Filtered-Soybean-Media.png\"><img loading=\"lazy\" decoding=\"async\" class=\"alignnone wp-image-1010\" src=\"https:\/\/baliga.systemsbiology.net\/see-interns\/hs2016\/wp-content\/uploads\/sites\/4\/2016\/08\/Filtered-Soybean-Media-300x167.png\" alt=\"Filtered Soybean Media\" width=\"490\" height=\"273\" srcset=\"https:\/\/baliga.systemsbiology.net\/see-interns\/hs2016\/wp-content\/uploads\/sites\/4\/2016\/08\/Filtered-Soybean-Media-300x167.png 300w, https:\/\/baliga.systemsbiology.net\/see-interns\/hs2016\/wp-content\/uploads\/sites\/4\/2016\/08\/Filtered-Soybean-Media-768x427.png 768w, https:\/\/baliga.systemsbiology.net\/see-interns\/hs2016\/wp-content\/uploads\/sites\/4\/2016\/08\/Filtered-Soybean-Media.png 910w\" sizes=\"auto, (max-width: 490px) 100vw, 490px\" \/><\/a><\/p>\n<p>Filtered soybeans work perfectly for <em>Halobacterium<\/em>. Halo showed a beautiful growth pattern.<\/p>\n<p><a href=\"https:\/\/baliga.systemsbiology.net\/see-interns\/hs2016\/wp-content\/uploads\/sites\/4\/2016\/08\/Unfiltered-Soybean-Media-1.png\"><img loading=\"lazy\" decoding=\"async\" class=\"alignnone wp-image-1011\" src=\"https:\/\/baliga.systemsbiology.net\/see-interns\/hs2016\/wp-content\/uploads\/sites\/4\/2016\/08\/Unfiltered-Soybean-Media-1-300x167.png\" alt=\"Unfiltered Soybean Media\" width=\"490\" height=\"273\" srcset=\"https:\/\/baliga.systemsbiology.net\/see-interns\/hs2016\/wp-content\/uploads\/sites\/4\/2016\/08\/Unfiltered-Soybean-Media-1-300x167.png 300w, https:\/\/baliga.systemsbiology.net\/see-interns\/hs2016\/wp-content\/uploads\/sites\/4\/2016\/08\/Unfiltered-Soybean-Media-1-768x428.png 768w, https:\/\/baliga.systemsbiology.net\/see-interns\/hs2016\/wp-content\/uploads\/sites\/4\/2016\/08\/Unfiltered-Soybean-Media-1.png 909w\" sizes=\"auto, (max-width: 490px) 100vw, 490px\" \/><\/a><\/p>\n<p>Unfiltered soybeans do not work. Large chunks of soybean interfered with the optical densities.<\/p>\n<p><a href=\"https:\/\/baliga.systemsbiology.net\/see-interns\/hs2016\/wp-content\/uploads\/sites\/4\/2016\/08\/Whey-Media.png\"><img loading=\"lazy\" decoding=\"async\" class=\"alignnone wp-image-1012\" src=\"https:\/\/baliga.systemsbiology.net\/see-interns\/hs2016\/wp-content\/uploads\/sites\/4\/2016\/08\/Whey-Media-300x167.png\" alt=\"Whey Media\" width=\"490\" height=\"273\" srcset=\"https:\/\/baliga.systemsbiology.net\/see-interns\/hs2016\/wp-content\/uploads\/sites\/4\/2016\/08\/Whey-Media-300x167.png 300w, https:\/\/baliga.systemsbiology.net\/see-interns\/hs2016\/wp-content\/uploads\/sites\/4\/2016\/08\/Whey-Media-768x428.png 768w, https:\/\/baliga.systemsbiology.net\/see-interns\/hs2016\/wp-content\/uploads\/sites\/4\/2016\/08\/Whey-Media.png 909w\" sizes=\"auto, (max-width: 490px) 100vw, 490px\" \/><\/a><\/p>\n<p style=\"text-align: justify\">This graph compares whey media isolate and whey protein powder. Both show an increase from original values but have many dips along the way. The data was inconsistent and thus unreliable.<\/p>\n<p><a href=\"https:\/\/baliga.systemsbiology.net\/see-interns\/hs2016\/wp-content\/uploads\/sites\/4\/2016\/08\/Epsom-Salt.png\"><img loading=\"lazy\" decoding=\"async\" class=\"alignnone wp-image-1013\" src=\"https:\/\/baliga.systemsbiology.net\/see-interns\/hs2016\/wp-content\/uploads\/sites\/4\/2016\/08\/Epsom-Salt-300x183.png\" alt=\"Epsom Salt\" width=\"490\" height=\"299\" srcset=\"https:\/\/baliga.systemsbiology.net\/see-interns\/hs2016\/wp-content\/uploads\/sites\/4\/2016\/08\/Epsom-Salt-300x183.png 300w, https:\/\/baliga.systemsbiology.net\/see-interns\/hs2016\/wp-content\/uploads\/sites\/4\/2016\/08\/Epsom-Salt-768x469.png 768w, https:\/\/baliga.systemsbiology.net\/see-interns\/hs2016\/wp-content\/uploads\/sites\/4\/2016\/08\/Epsom-Salt.png 909w\" sizes=\"auto, (max-width: 490px) 100vw, 490px\" \/><\/a><\/p>\n<p>Epsom Salt result in a steady increase of halo population and thus we can conclude that it works.<\/p>\n<p><a href=\"https:\/\/baliga.systemsbiology.net\/see-interns\/hs2016\/wp-content\/uploads\/sites\/4\/2016\/08\/Kosher-Salt.png\"><img loading=\"lazy\" decoding=\"async\" class=\"alignnone wp-image-1014\" src=\"https:\/\/baliga.systemsbiology.net\/see-interns\/hs2016\/wp-content\/uploads\/sites\/4\/2016\/08\/Kosher-Salt-300x183.png\" alt=\"Kosher Salt\" width=\"490\" height=\"299\" srcset=\"https:\/\/baliga.systemsbiology.net\/see-interns\/hs2016\/wp-content\/uploads\/sites\/4\/2016\/08\/Kosher-Salt-300x183.png 300w, https:\/\/baliga.systemsbiology.net\/see-interns\/hs2016\/wp-content\/uploads\/sites\/4\/2016\/08\/Kosher-Salt-768x469.png 768w, https:\/\/baliga.systemsbiology.net\/see-interns\/hs2016\/wp-content\/uploads\/sites\/4\/2016\/08\/Kosher-Salt.png 909w\" sizes=\"auto, (max-width: 490px) 100vw, 490px\" \/><\/a><\/p>\n<p>Kosher Salt gave us beautiful results. The growth curve shows an increase in halo.<\/p>\n<h4>Future Work<\/h4>\n<p style=\"text-align: justify\">In the future we hope to find substitutes for sodium citrate as well as conduct the experiments with whey media again after filtering the media to get better results.<\/p>\n<p>&nbsp;<\/p>\n<p>&nbsp;<\/p>\n<p>&nbsp;<\/p>\n","protected":false},"excerpt":{"rendered":"<p>By Sumaiya Sathar Introduction Halobacterium salinarum is a type of archaea that resides in highly salty environments. H. salinarum is widely used as a model organism since their genetic processing method is very similar to that of humans. In a laboratory setting, halobacterium is grown in Complete Media which is a combination of laboratory-grade chemicals and proteins that provide the nutrients H. salinarum needs in order to survive.\u00a0Halobacterium is extremely simple to conduct experiments with and safely handle, thus making it a favorite amongst teachers&hellip;<\/p>\n","protected":false},"author":14,"featured_media":0,"parent":193,"menu_order":0,"comment_status":"closed","ping_status":"closed","template":"","meta":{"footnotes":""},"class_list":["post-176","page","type-page","status-publish","hentry"],"_links":{"self":[{"href":"https:\/\/baliga.systemsbiology.net\/see-interns\/hs2016\/wp-json\/wp\/v2\/pages\/176","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/baliga.systemsbiology.net\/see-interns\/hs2016\/wp-json\/wp\/v2\/pages"}],"about":[{"href":"https:\/\/baliga.systemsbiology.net\/see-interns\/hs2016\/wp-json\/wp\/v2\/types\/page"}],"author":[{"embeddable":true,"href":"https:\/\/baliga.systemsbiology.net\/see-interns\/hs2016\/wp-json\/wp\/v2\/users\/14"}],"replies":[{"embeddable":true,"href":"https:\/\/baliga.systemsbiology.net\/see-interns\/hs2016\/wp-json\/wp\/v2\/comments?post=176"}],"version-history":[{"count":12,"href":"https:\/\/baliga.systemsbiology.net\/see-interns\/hs2016\/wp-json\/wp\/v2\/pages\/176\/revisions"}],"predecessor-version":[{"id":1017,"href":"https:\/\/baliga.systemsbiology.net\/see-interns\/hs2016\/wp-json\/wp\/v2\/pages\/176\/revisions\/1017"}],"up":[{"embeddable":true,"href":"https:\/\/baliga.systemsbiology.net\/see-interns\/hs2016\/wp-json\/wp\/v2\/pages\/193"}],"wp:attachment":[{"href":"https:\/\/baliga.systemsbiology.net\/see-interns\/hs2016\/wp-json\/wp\/v2\/media?parent=176"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}