{"id":410,"date":"2016-06-15T18:43:22","date_gmt":"2016-06-16T00:43:22","guid":{"rendered":"http:\/\/ciresblogs.colorado.edu\/reccs\/?p=410"},"modified":"2016-06-15T18:55:43","modified_gmt":"2016-06-16T00:55:43","slug":"extracting-peptidoglycan-fragments","status":"publish","type":"post","link":"https:\/\/ciresblogs.colorado.edu\/reccs\/2016\/06\/15\/extracting-peptidoglycan-fragments\/","title":{"rendered":"Extracting Peptidoglycan Fragments"},"content":{"rendered":"<p><strong>June 9<sup>th<\/sup>, 2016<\/strong><\/p>\n<p>I showed up to the lab happy to find out that the peptidoglycan extraction hadn\u2019t happened yet! I was excited that I would be able to see how to go from having a concentrated solution of bacteria cells to only having fragments of their peptidoglycan leftover.<\/p>\n<p><a href=\"http:\/\/ciresblogs.colorado.edu\/reccs\/wp-content\/uploads\/sites\/46\/2016\/06\/Addition-of-10-TCA-into-Bacteria-Pellet-6-9-16-e1466037564548.jpg\"><img loading=\"lazy\" decoding=\"async\" class=\" wp-image-411 alignleft\" src=\"http:\/\/ciresblogs.colorado.edu\/reccs\/wp-content\/uploads\/sites\/46\/2016\/06\/Addition-of-10-TCA-into-Bacteria-Pellet-6-9-16-e1466037564548-169x300.jpg\" alt=\"Addition of 10% TCA into Bacteria Pellet [6-9-16]\" width=\"148\" height=\"263\" srcset=\"https:\/\/ciresblogs.colorado.edu\/reccs\/wp-content\/uploads\/sites\/46\/2016\/06\/Addition-of-10-TCA-into-Bacteria-Pellet-6-9-16-e1466037564548-169x300.jpg 169w, https:\/\/ciresblogs.colorado.edu\/reccs\/wp-content\/uploads\/sites\/46\/2016\/06\/Addition-of-10-TCA-into-Bacteria-Pellet-6-9-16-e1466037564548-768x1365.jpg 768w, https:\/\/ciresblogs.colorado.edu\/reccs\/wp-content\/uploads\/sites\/46\/2016\/06\/Addition-of-10-TCA-into-Bacteria-Pellet-6-9-16-e1466037564548-576x1024.jpg 576w, https:\/\/ciresblogs.colorado.edu\/reccs\/wp-content\/uploads\/sites\/46\/2016\/06\/Addition-of-10-TCA-into-Bacteria-Pellet-6-9-16-e1466037564548.jpg 1836w\" sizes=\"auto, (max-width: 148px) 100vw, 148px\" \/><\/a>Although I was ecstatic for learning the process, I was a bit saddened that I was not able to help with the extraction at all. I walked around the lab following Tess and Paul, another laboratory technician, like a duckling following its mother. I still made a point to ask questions when I didn\u2019t understand what was happening, or if I wasn\u2019t sure what was next in the procedure, but I didn\u2019t have the chance to touch or use any laboratory equipment at all. It was an interesting process to observe, nonetheless; the bacteria cells were killed with the addition of trichloroacetic acid and by boiling them in<a href=\"http:\/\/ciresblogs.colorado.edu\/reccs\/wp-content\/uploads\/sites\/46\/2016\/06\/Trichloroacetic-Acid-6-9-16-e1466037717471.jpg\"><img loading=\"lazy\" decoding=\"async\" class=\" wp-image-413 alignright\" src=\"http:\/\/ciresblogs.colorado.edu\/reccs\/wp-content\/uploads\/sites\/46\/2016\/06\/Trichloroacetic-Acid-6-9-16-e1466037717471-169x300.jpg\" alt=\"Trichloroacetic Acid [6-9-16]\" width=\"159\" height=\"282\" srcset=\"https:\/\/ciresblogs.colorado.edu\/reccs\/wp-content\/uploads\/sites\/46\/2016\/06\/Trichloroacetic-Acid-6-9-16-e1466037717471-169x300.jpg 169w, https:\/\/ciresblogs.colorado.edu\/reccs\/wp-content\/uploads\/sites\/46\/2016\/06\/Trichloroacetic-Acid-6-9-16-e1466037717471-768x1365.jpg 768w, https:\/\/ciresblogs.colorado.edu\/reccs\/wp-content\/uploads\/sites\/46\/2016\/06\/Trichloroacetic-Acid-6-9-16-e1466037717471-576x1024.jpg 576w, https:\/\/ciresblogs.colorado.edu\/reccs\/wp-content\/uploads\/sites\/46\/2016\/06\/Trichloroacetic-Acid-6-9-16-e1466037717471.jpg 1836w\" sizes=\"auto, (max-width: 159px) 100vw, 159px\" \/><\/a> glass tubes submerged\u00a0in a water bath \u2013 this ensured that the cells burst and their innards spilled out into a glob at the bottom of the tube, leftover DNA was digested with the addition of DNase and proteins were digested with the addition of trypsin. All that was left of the bacteria was some peptidoglycan fragments and peptides.<\/p>\n<p>Although I wasn\u2019t able to do any lab work, one particularly useful part of today was sitting down with Tess and scheduling a work plan. We discussed what was left to do for the experiment and set reasonable due dates for completing each task. We then made sure to keep a few weeks in July freed up just in case the experiment doesn\u2019t go exactly as planned. The last few weeks of my time in the laboratory will be spend processing data and prepping for the poster session and presentation.<\/p>\n<p>Part of <a href=\"http:\/\/ciresblogs.colorado.edu\/reccs\/wp-content\/uploads\/sites\/46\/2016\/06\/Weighing-of-Wet-Pellet-Samples-6-9-16-e1466037641596.jpg\"><img loading=\"lazy\" decoding=\"async\" class=\" wp-image-412 alignleft\" src=\"http:\/\/ciresblogs.colorado.edu\/reccs\/wp-content\/uploads\/sites\/46\/2016\/06\/Weighing-of-Wet-Pellet-Samples-6-9-16-e1466037641596-169x300.jpg\" alt=\"Weighing of Wet Pellet Samples [6-9-16]\" width=\"159\" height=\"283\" srcset=\"https:\/\/ciresblogs.colorado.edu\/reccs\/wp-content\/uploads\/sites\/46\/2016\/06\/Weighing-of-Wet-Pellet-Samples-6-9-16-e1466037641596-169x300.jpg 169w, https:\/\/ciresblogs.colorado.edu\/reccs\/wp-content\/uploads\/sites\/46\/2016\/06\/Weighing-of-Wet-Pellet-Samples-6-9-16-e1466037641596-768x1365.jpg 768w, https:\/\/ciresblogs.colorado.edu\/reccs\/wp-content\/uploads\/sites\/46\/2016\/06\/Weighing-of-Wet-Pellet-Samples-6-9-16-e1466037641596-576x1024.jpg 576w, https:\/\/ciresblogs.colorado.edu\/reccs\/wp-content\/uploads\/sites\/46\/2016\/06\/Weighing-of-Wet-Pellet-Samples-6-9-16-e1466037641596.jpg 1836w\" sizes=\"auto, (max-width: 159px) 100vw, 159px\" \/><\/a>our experiment is to subject the soil bacteria to 5 different resuscitation substances; substances that cause dormant bacteria cells to \u201cwake-up\u201d and start actively growing again. We\u2019ve already determined three that we can test: the peptidoglycan fragments, N-acyl homoserine lactones (AHLs), and volatile organic compounds (VOCs) from leaf litter. Tess assigned me to come up with 2 more by tomorrow \u2013 if the substance requires some sort of extraction procedure, and especially by Monday when we need to start testing the resuscitation substances. The pressure is on! I\u2019m feeling a little lost since I don\u2019t know what to look for and my understanding of the physiology of microbes and the availability of supplies in the lab is painfully novice. I\u2019m hoping that I can come up with something\u2026 I will definitely read some articles and look through some textbooks to hopefully have 2 ideas for substances to try that have a relatively reasonable validation behind choosing them.<\/p>\n","protected":false},"excerpt":{"rendered":"<p>June 9th, 2016 I showed up to the lab happy to find out that the peptidoglycan extraction hadn\u2019t happened yet! I was excited that I would be able to see how to go from having a concentrated solution of bacteria cells to only having fragments of their peptidoglycan leftover. Although I was ecstatic for learning&hellip; <a class=\"read-more\" href=\"https:\/\/ciresblogs.colorado.edu\/reccs\/2016\/06\/15\/extracting-peptidoglycan-fragments\/\">Read More<\/a><\/p>\n","protected":false},"author":79,"featured_media":0,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[4],"tags":[],"class_list":["post-410","post","type-post","status-publish","format-standard","hentry","category-reccs-2016"],"jetpack_featured_media_url":"","publishpress_future_action":{"enabled":false,"date":"2026-08-29 21:11:31","action":"change-status","newStatus":"draft","terms":[],"taxonomy":"category","extraData":[]},"publishpress_future_workflow_manual_trigger":{"enabledWorkflows":[]},"_links":{"self":[{"href":"https:\/\/ciresblogs.colorado.edu\/reccs\/wp-json\/wp\/v2\/posts\/410","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/ciresblogs.colorado.edu\/reccs\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/ciresblogs.colorado.edu\/reccs\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/ciresblogs.colorado.edu\/reccs\/wp-json\/wp\/v2\/users\/79"}],"replies":[{"embeddable":true,"href":"https:\/\/ciresblogs.colorado.edu\/reccs\/wp-json\/wp\/v2\/comments?post=410"}],"version-history":[{"count":1,"href":"https:\/\/ciresblogs.colorado.edu\/reccs\/wp-json\/wp\/v2\/posts\/410\/revisions"}],"predecessor-version":[{"id":414,"href":"https:\/\/ciresblogs.colorado.edu\/reccs\/wp-json\/wp\/v2\/posts\/410\/revisions\/414"}],"wp:attachment":[{"href":"https:\/\/ciresblogs.colorado.edu\/reccs\/wp-json\/wp\/v2\/media?parent=410"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/ciresblogs.colorado.edu\/reccs\/wp-json\/wp\/v2\/categories?post=410"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/ciresblogs.colorado.edu\/reccs\/wp-json\/wp\/v2\/tags?post=410"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}