{"id":861,"date":"2013-06-18T11:18:34","date_gmt":"2013-06-18T16:18:34","guid":{"rendered":"http:\/\/blogs.acu.edu\/biologyresearch\/?p=861"},"modified":"2013-07-13T11:19:55","modified_gmt":"2013-07-13T16:19:55","slug":"week-1","status":"publish","type":"post","link":"https:\/\/blogs.acu.edu\/biologyresearch\/2013\/06\/18\/week-1\/","title":{"rendered":"Week 1"},"content":{"rendered":"

\"Davis<\/a>First week back in the CS lab!<\/p>\n

I actually started back in the lab on May 29th and, go figure, Baby Huddleston decided he was ready to meet his family! Of course, Dr. Huddleston, who was\u00a0in labor<\/em>, was worried about running a gel on a vector she made and asked me to do it for her. That lady. I was more than happy to do it for her! It felt so good to be back in the lab!\u00a0I really missed it in there. It was so nice because nobody else was in the lab so I shut the doors and played my music loud. I started initial PCRs for my SOE PCR that week. I got 2 out of my 3 of my needed gene combo products done!<\/p>\n

So, the official first week back in the lab and there\u2019s a lot of new changes! First, meeting at 8 am (really bad since I\u2019d been waking up at 4pm every other day). Second, I had two new lab partners (since Kristin and Zach left me to be adults and go to dental and medical school. Whatever.). \u00a0Third, Dr. H was out being a good mom. And fourth, \u00a0I was bestowed the honor of showing the new kids the way around the lab. They\u2019re Madeline, my dear friend who lived across the hall from me freshman year, and Jacob, a\u00a0freshman<\/del>\u00a0sophomore who is actually the sweetest. I am so excited to spend the summer with them.<\/p>\n

We\u2019re beginning the summer with SOE\u00a0(splice overlap extension)\u00a0PCR . I am so excited because we\u2019ve been talking about doing SOE for over a year and we\u2019re finally here! On our agenda this week we needed to do the initial PCR of their products so we can sew (get it??) them together. We\u2019re replacing our genes with a gentamicin cassette to be used for screening. It\u2019s sort of bizarre teaching people how to get around the lab because it\u2019s second nature to me now. They were asking me how to pipette product into the wells in a gel, and I stood there for a minute just going\u2026 uhhhhh. I had to watch myself do my technique so I could articulate it to them. Funny how far I\u2019ve come. I was in their shoes a year ago. Poor guys are stuck with me and not Dr. H!<\/p>\n

Once we obtained and confirmed our initial products, they were purified to be used for the second step of SOE PCR: the first SOE reaction. This will bind initial products A and B to each other. Both Madeline and I encountered the same problem: we obtained products AB of the correct size, 1kb, but we also had products that were 500bp, which indicated to me that the pieces of DNA were breaking up into their initial products, A and B, or they weren\u2019t binding together at all and the gel was picking up the original amount of template added to the PCR reaction (thanks Dr. Brokaw).<\/p>\n

So, the plan was to change the annealing temperatures and extension times to see if we obtained better results for the next week. Until\u2026<\/p>\n","protected":false},"excerpt":{"rendered":"

First week back in the CS lab! I actually started back in the lab on May 29th and, go figure, Baby Huddleston decided he was ready to meet his family! Of course, Dr. Huddleston, who was\u00a0in labor, was worried about … Continue reading →<\/span><\/a><\/p>\n","protected":false},"author":5523,"featured_media":0,"comment_status":"open","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"_et_pb_use_builder":"","_et_pb_old_content":"","_et_gb_content_width":"","_monsterinsights_skip_tracking":false,"_monsterinsights_sitenote_active":false,"_monsterinsights_sitenote_note":"","_monsterinsights_sitenote_category":0,"footnotes":""},"categories":[55710],"tags":[55715,55717,55719,55724],"class_list":["post-861","post","type-post","status-publish","format-standard","hentry","category-huddleston-lab","tag-dna","tag-genetics","tag-microbiology","tag-molecular-biology"],"_links":{"self":[{"href":"https:\/\/blogs.acu.edu\/biologyresearch\/wp-json\/wp\/v2\/posts\/861","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/blogs.acu.edu\/biologyresearch\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/blogs.acu.edu\/biologyresearch\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/blogs.acu.edu\/biologyresearch\/wp-json\/wp\/v2\/users\/5523"}],"replies":[{"embeddable":true,"href":"https:\/\/blogs.acu.edu\/biologyresearch\/wp-json\/wp\/v2\/comments?post=861"}],"version-history":[{"count":2,"href":"https:\/\/blogs.acu.edu\/biologyresearch\/wp-json\/wp\/v2\/posts\/861\/revisions"}],"predecessor-version":[{"id":864,"href":"https:\/\/blogs.acu.edu\/biologyresearch\/wp-json\/wp\/v2\/posts\/861\/revisions\/864"}],"wp:attachment":[{"href":"https:\/\/blogs.acu.edu\/biologyresearch\/wp-json\/wp\/v2\/media?parent=861"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/blogs.acu.edu\/biologyresearch\/wp-json\/wp\/v2\/categories?post=861"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/blogs.acu.edu\/biologyresearch\/wp-json\/wp\/v2\/tags?post=861"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}