{"id":290,"date":"2014-10-16T18:30:31","date_gmt":"2014-10-16T18:30:31","guid":{"rendered":"https:\/\/www.bates.edu\/williams-lab\/?p=290"},"modified":"2024-01-04T16:39:20","modified_gmt":"2024-01-04T21:39:20","slug":"6-weeks-into-thesis","status":"publish","type":"post","link":"https:\/\/www.bates.edu\/williams-lab\/2014\/10\/16\/6-weeks-into-thesis\/","title":{"rendered":"6 weeks into thesis!"},"content":{"rendered":"<p>We are now 6 weeks into thesis!<\/p>\n<p>It certainly hasn\u2019t felt like 6 weeks. We\u2019re wrapping up a bunch of prerequisites before we start our experimental trials. These include primer efficiency curves, as well as making sure our housekeeping genes are good picks for our chemical doses. We will be comparing expression of housekeeping genes in dosed and non-dosed embryos in order to see if expression really is being changed. As a reminder, we are looking at the expression of six antioxidant genes that have been implicated in the oxidative stress response and with our four chemicals, tBOOH, tBHQ, Diquat, and sulforphane. Lab has been great, breeding our fish is getting easier and faster, and I\u2019ve gotten much more comfortable with the procedures we\u2019ve been doing over the past few weeks. I\u2019m still a little wary of RNA extractions, however I feel confident I can do them. It\u2019s pretty nerve wracking considering that if we do mess up, we have to start all over again. To get a sample at this point, we have to breed for embryos, collect the embryos, dosed them with chemical, and froze the embryos.<br \/>\n<div id=\"attachment_291\" style=\"width: 310px\" class=\"wp-caption alignnone\"><a href=\"https:\/\/www.bates.edu\/williams-lab\/files\/2014\/10\/ZF-breeding-1.png\"><img loading=\"lazy\" decoding=\"async\" aria-describedby=\"caption-attachment-291\" src=\"https:\/\/www.bates.edu\/williams-lab\/files\/2014\/10\/ZF-breeding-1-300x81.png\" alt=\"Breeding zebrafish for embryos\" width=\"300\" height=\"81\" class=\"size-medium wp-image-291\" srcset=\"https:\/\/www.bates.edu\/williams-lab\/files\/2014\/10\/ZF-breeding-1-300x81.png 300w, https:\/\/www.bates.edu\/williams-lab\/files\/2014\/10\/ZF-breeding-1-620x168.png 620w, https:\/\/www.bates.edu\/williams-lab\/files\/2014\/10\/ZF-breeding-1.png 640w\" sizes=\"(max-width: 300px) 100vw, 300px\" \/><\/a><p id=\"caption-attachment-291\" class=\"wp-caption-text\">Breeding zebrafish for embryos<\/p><\/div><br \/>\nAfter that, we have to extract the RNA from those embryos then finally run qPCR with our housekeeping and antioxidant genes. If you mess up at the RNA extraction phase, you have to breed and dose all over again! So, while it is a bit of pressure, it\u2019s certainly doable. Experimental trials will add an extra step of microinjection of morpholino to knockdown Nfe2, which I will explain further in my next blog post. I\u2019ve also delved into writing my actual thesis, and I have to say I can\u2019t sing the praises of summer research enough. It gave me a huge head start on sources and introductory knowledge, and it\u2019s been nice getting to dive deeper and deeper into the primary literature given that I\u2019ve got a good background of our topic. It only helps that I personally find ROS chemistry pretty fascinating. With October coming to a close, I can\u2019t wait to start experimental trials soon!<\/p>\n","protected":false},"excerpt":{"rendered":"<p>We are now 6 weeks into thesis! It certainly hasn\u2019t felt like&hellip;<\/p>\n","protected":false},"author":381,"featured_media":291,"comment_status":"closed","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"_hide_ai_chatbot":false,"_ai_chatbot_style":"","associated_faculty":[],"_Page_Specific_Css":"","_bates_restrict_mod":false,"_batesModPostContentOverride_prepend":false,"_batesModPostContentOverride_append":false,"_batesModPostContentOverride_append_before_footer":false,"_table_of_contents_display":false,"_table_of_contents_location":"","_table_of_contents_disableSticky":false,"_is_featured":false,"footnotes":"","_bates_seo_meta_description":"","_bates_seo_block_robots":false,"_bates_seo_sharing_image_id":0,"_bates_seo_sharing_image_twitter_id":0,"_bates_seo_share_title":"","_bates_seo_canonical_overwrite":"","_bates_seo_twitter_template":""},"categories":[1],"tags":[],"class_list":["post-290","post","type-post","status-publish","format-standard","has-post-thumbnail","hentry","category-blog"],"_links":{"self":[{"href":"https:\/\/www.bates.edu\/williams-lab\/wp-json\/wp\/v2\/posts\/290","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/www.bates.edu\/williams-lab\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/www.bates.edu\/williams-lab\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/www.bates.edu\/williams-lab\/wp-json\/wp\/v2\/users\/381"}],"replies":[{"embeddable":true,"href":"https:\/\/www.bates.edu\/williams-lab\/wp-json\/wp\/v2\/comments?post=290"}],"version-history":[{"count":1,"href":"https:\/\/www.bates.edu\/williams-lab\/wp-json\/wp\/v2\/posts\/290\/revisions"}],"predecessor-version":[{"id":292,"href":"https:\/\/www.bates.edu\/williams-lab\/wp-json\/wp\/v2\/posts\/290\/revisions\/292"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/www.bates.edu\/williams-lab\/wp-json\/wp\/v2\/media\/291"}],"wp:attachment":[{"href":"https:\/\/www.bates.edu\/williams-lab\/wp-json\/wp\/v2\/media?parent=290"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/www.bates.edu\/williams-lab\/wp-json\/wp\/v2\/categories?post=290"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/www.bates.edu\/williams-lab\/wp-json\/wp\/v2\/tags?post=290"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}