{"id":115,"date":"2018-06-13T11:50:57","date_gmt":"2018-06-13T15:50:57","guid":{"rendered":"https:\/\/wordpress-projects.wolfware.ncsu.edu\/bio-183-lab-yhnn8gz\/?page_id=115"},"modified":"2020-04-15T15:06:14","modified_gmt":"2020-04-15T19:06:14","slug":"unit-4-lab-2-3-gene-transformation","status":"publish","type":"page","link":"https:\/\/wordpress-projects.wolfware.ncsu.edu\/bio-183-lab-yhnn8gz\/unit-4-lab-2-3-gene-transformation\/","title":{"rendered":"Unit 4: Labs 2 &amp; 3"},"content":{"rendered":"<h2><a href=\"https:\/\/wordpress-projects.wolfware.ncsu.edu\/bio-183-lab-yhnn8gz\/wp-content\/uploads\/sites\/74\/2018\/05\/GFP-jellyfish.jpg\"><img loading=\"lazy\" decoding=\"async\" class=\"alignnone size-thumbnail wp-image-66\" src=\"https:\/\/wordpress-projects.wolfware.ncsu.edu\/bio-183-lab-yhnn8gz\/wp-content\/uploads\/sites\/74\/2018\/05\/GFP-jellyfish-150x150.jpg\" alt=\"Glowing jellyfish\" width=\"150\" height=\"150\" \/><\/a><\/h2>\n<h2>Gene Transformation (2 weeks of lab)<\/h2>\n<p>&nbsp;<\/p>\n<p><a href=\"https:\/\/wordpress-projects.wolfware.ncsu.edu\/bio-183-lab-yhnn8gz\/wp-content\/uploads\/sites\/74\/2018\/06\/images.jpg\"><img loading=\"lazy\" decoding=\"async\" class=\"alignnone size-full wp-image-312\" src=\"https:\/\/wordpress-projects.wolfware.ncsu.edu\/bio-183-lab-yhnn8gz\/wp-content\/uploads\/sites\/74\/2018\/06\/images.jpg\" alt=\"pGlo Plasmid DNA\" width=\"225\" height=\"224\" srcset=\"https:\/\/wordpress-projects.wolfware.ncsu.edu\/bio-183-lab-yhnn8gz\/wp-content\/uploads\/sites\/74\/2018\/06\/images.jpg 225w, https:\/\/wordpress-projects.wolfware.ncsu.edu\/bio-183-lab-yhnn8gz\/wp-content\/uploads\/sites\/74\/2018\/06\/images-150x150.jpg 150w\" sizes=\"auto, (max-width: 225px) 100vw, 225px\" \/><\/a>\u00a0<a href=\"https:\/\/wordpress-projects.wolfware.ncsu.edu\/bio-183-lab-yhnn8gz\/wp-content\/uploads\/sites\/74\/2018\/06\/pGLO_Circle.jpg\"><img loading=\"lazy\" decoding=\"async\" class=\"alignnone wp-image-313 size-full\" src=\"https:\/\/wordpress-projects.wolfware.ncsu.edu\/bio-183-lab-yhnn8gz\/wp-content\/uploads\/sites\/74\/2018\/06\/pGLO_Circle.jpg\" alt=\"pGlo plasmid and transformed bacterial culture\" width=\"457\" height=\"154\" srcset=\"https:\/\/wordpress-projects.wolfware.ncsu.edu\/bio-183-lab-yhnn8gz\/wp-content\/uploads\/sites\/74\/2018\/06\/pGLO_Circle.jpg 457w, https:\/\/wordpress-projects.wolfware.ncsu.edu\/bio-183-lab-yhnn8gz\/wp-content\/uploads\/sites\/74\/2018\/06\/pGLO_Circle-300x101.jpg 300w\" sizes=\"auto, (max-width: 457px) 100vw, 457px\" \/><\/a><\/p>\n<p>&nbsp;<\/p>\n<h2>Week 1<\/h2>\n<h3><span style=\"color: #000000\">Background:<\/span><\/h3>\n<h3><a class=\"style11\" href=\"http:\/\/www.youtube.com\/watch?v=Sl2PRHGpYuU&amp;feature=related\">How GFP is used in research<\/a><span class=\"style11\">\u00a0<span style=\"color: #000000\">(10:03)<\/span><\/span><\/h3>\n<h3><span class=\"style11\"><a href=\"http:\/\/www.youtube.com\/watch?v=GmhD-RWNL6c&amp;sns=fb\">Funding Basic Science to Revolutionize Medicine<\/a>\u00a0<span style=\"color: #000000\">(3:59) that show how the focus in research has changed over the past 50 years.<\/span><\/span><\/h3>\n<h3><span class=\"style11\"><a href=\"https:\/\/news.stanford.edu\/news\/2015\/september\/plants-drug-sattely-091015.html\">Stanford Scientists Produce Cancer Drugs from Rare Plants<\/a>\u00a0<span style=\"color: #000000\">(1:59) by making plasmids and inserting them into more easily grown plants or yeasts.<\/span><\/span><\/h3>\n<p>&nbsp;<\/p>\n<h3><span style=\"color: #000000\">Important Concepts in the Transformation Procedures:<\/span><\/h3>\n<h3><a href=\"https:\/\/www.youtube.com\/watch?v=Ik_Pxht1LM0\">Using Restriction Enzymes and DNA Ligase to make plasmids<\/a>\u00a0<span style=\"color: #000000\">(1:38)<\/span><\/h3>\n<h3><span class=\"style11\"><a href=\"https:\/\/wordpress-projects.wolfware.ncsu.edu\/bio-183-lab-yhnn8gz\/wp-content\/uploads\/sites\/74\/2018\/06\/pGlo-Plasmid.pdf\" target=\"_blank\" rel=\"noopener noreferrer\">pGlo Plasmid<\/a> <span style=\"color: #000000\">we will use in lab.<\/span><\/span><\/h3>\n<h3><span class=\"style11\"><a href=\"http:\/\/dnalc.org\/resources\/animations\/transformation2.html\">DNA transformation using plasmids<\/a>\u00a0<span style=\"color: #000000\">&#8211; animation explaining the steps in our Transformation procedure.\u00a0 It is very important to understand the nature of the bacterial cell wall, why CaCl<sub>2<\/sub> is used, and why the heat shock is important.<\/span><\/span><\/h3>\n<h3><a href=\"https:\/\/www.youtube.com\/watch?v=c40UudFIlGw\" target=\"_blank\" rel=\"noopener noreferrer\">Video of BIORAD pGlo Transformation procedure (p. 23-28 in your lab manual)<\/a><\/h3>\n<p>&nbsp;<\/p>\n<p><strong>Measurement marks on disposable plastic pipettes provided in the BIORAD kit:<\/strong><\/p>\n<p><a href=\"https:\/\/wordpress-projects.wolfware.ncsu.edu\/bio-183-lab-yhnn8gz\/wp-content\/uploads\/sites\/74\/2018\/06\/1-ml-pipet-diagram.jpg\"><img loading=\"lazy\" decoding=\"async\" class=\"alignnone size-full wp-image-314\" src=\"https:\/\/wordpress-projects.wolfware.ncsu.edu\/bio-183-lab-yhnn8gz\/wp-content\/uploads\/sites\/74\/2018\/06\/1-ml-pipet-diagram.jpg\" alt=\"pipet measurements\" width=\"720\" height=\"540\" srcset=\"https:\/\/wordpress-projects.wolfware.ncsu.edu\/bio-183-lab-yhnn8gz\/wp-content\/uploads\/sites\/74\/2018\/06\/1-ml-pipet-diagram.jpg 720w, https:\/\/wordpress-projects.wolfware.ncsu.edu\/bio-183-lab-yhnn8gz\/wp-content\/uploads\/sites\/74\/2018\/06\/1-ml-pipet-diagram-300x225.jpg 300w\" sizes=\"auto, (max-width: 720px) 100vw, 720px\" \/><\/a><\/p>\n<p>&nbsp;<\/p>\n<h3>Week 1 &#8211; Hypotheses for Results of each of the agar plates inoculated:<\/h3>\n<ul>\n<li><span style=\"color: #000000;font-size: 12pt\">Fill out the table on page 29 to indicate what you think the results would be for each of the 4 agar plates that were inoculated in this procedure.\u00a0 Also fill out the gray boxes for the agar plates that were not inoculated.\u00a0 Include why you think you&#8217;d get these results in ALL the boxes as well.<\/span><\/li>\n<\/ul>\n<p>&nbsp;<\/p>\n<h2>Week 2<\/h2>\n<h3><span style=\"color: #000000\">Actual Results:\u00a0 The following pictures show the bacterial growth on the different nutrient agar plates under white light and UV light from a previous semester.<\/span><\/h3>\n<ul>\n<li><span style=\"font-size: 12pt\">Record the data in Table 1 page 33 that corresponds to the Group\/Table where you normally sit in the lab room.\u00a0 Be sure to collect the data from both the White Light and the UV Light pictures.\u00a0 For example, if you sit at Table 3 in the room, you only need to record data from the Group 3 pictures &#8211; both white light (3-WL) and UV light (3-UV).<\/span><\/li>\n<li><span style=\"font-size: 12pt\">Answer the questions on pages 33-34.\u00a0<\/span><\/li>\n<\/ul>\n<p>&nbsp;<\/p>\n<table style=\"border-collapse: collapse;width: 100%\">\n<tbody>\n<tr>\n<td style=\"width: 14.2857%\"><strong>Light Source<\/strong><\/td>\n<td style=\"width: 14.2857%\"><strong>Group 1<\/strong><\/td>\n<td style=\"width: 14.2857%\"><strong>Group 2<\/strong><\/td>\n<td style=\"width: 14.2857%\"><strong>Group 3<\/strong><\/td>\n<td style=\"width: 14.2857%\"><strong>Group 4<\/strong><\/td>\n<td style=\"width: 14.2857%\"><strong>Group 5<\/strong><\/td>\n<td style=\"width: 14.2857%\"><strong>Group 6<\/strong><\/td>\n<\/tr>\n<tr>\n<td style=\"width: 14.2857%\"><strong>White Light (WL)<\/strong><\/td>\n<td style=\"width: 14.2857%\"><a href=\"https:\/\/wordpress-projects.wolfware.ncsu.edu\/bio-183-lab-yhnn8gz\/wp-content\/uploads\/sites\/74\/2020\/03\/Set1.jpg\" target=\"_blank\" rel=\"noopener noreferrer\">1-WL<\/a><\/td>\n<td style=\"width: 14.2857%\"><a href=\"https:\/\/wordpress-projects.wolfware.ncsu.edu\/bio-183-lab-yhnn8gz\/wp-content\/uploads\/sites\/74\/2020\/03\/Set2.jpg\" target=\"_blank\" rel=\"noopener noreferrer\">2-WL<\/a><\/td>\n<td style=\"width: 14.2857%\"><a href=\"https:\/\/wordpress-projects.wolfware.ncsu.edu\/bio-183-lab-yhnn8gz\/wp-content\/uploads\/sites\/74\/2020\/03\/Set3.jpg\" target=\"_blank\" rel=\"noopener noreferrer\">3-WL<\/a><\/td>\n<td style=\"width: 14.2857%\"><a href=\"https:\/\/wordpress-projects.wolfware.ncsu.edu\/bio-183-lab-yhnn8gz\/wp-content\/uploads\/sites\/74\/2020\/03\/Set4-1.jpg\" target=\"_blank\" rel=\"noopener noreferrer\">4-WL<\/a><\/td>\n<td style=\"width: 14.2857%\"><a href=\"https:\/\/wordpress-projects.wolfware.ncsu.edu\/bio-183-lab-yhnn8gz\/wp-content\/uploads\/sites\/74\/2020\/03\/Set-5.jpg\" target=\"_blank\" rel=\"noopener noreferrer\">5-WL<\/a><\/td>\n<td style=\"width: 14.2857%\"><a href=\"https:\/\/wordpress-projects.wolfware.ncsu.edu\/bio-183-lab-yhnn8gz\/wp-content\/uploads\/sites\/74\/2020\/03\/Set6-1.jpg\" target=\"_blank\" rel=\"noopener noreferrer\">6-WL<\/a><\/td>\n<\/tr>\n<tr>\n<td style=\"width: 14.2857%\"><strong>UV Light (UV)<\/strong><\/td>\n<td style=\"width: 14.2857%\"><a href=\"https:\/\/wordpress-projects.wolfware.ncsu.edu\/bio-183-lab-yhnn8gz\/wp-content\/uploads\/sites\/74\/2020\/03\/Set1-1.jpg\" target=\"_blank\" rel=\"noopener noreferrer\">1-UV<\/a><\/td>\n<td style=\"width: 14.2857%\"><a href=\"https:\/\/wordpress-projects.wolfware.ncsu.edu\/bio-183-lab-yhnn8gz\/wp-content\/uploads\/sites\/74\/2020\/03\/Set2-1.jpg\" target=\"_blank\" rel=\"noopener noreferrer\">2-UV<\/a><\/td>\n<td style=\"width: 14.2857%\"><a href=\"https:\/\/wordpress-projects.wolfware.ncsu.edu\/bio-183-lab-yhnn8gz\/wp-content\/uploads\/sites\/74\/2020\/03\/Set3-1.jpg\" target=\"_blank\" rel=\"noopener noreferrer\">3-UV<\/a><\/td>\n<td style=\"width: 14.2857%\"><a href=\"https:\/\/wordpress-projects.wolfware.ncsu.edu\/bio-183-lab-yhnn8gz\/wp-content\/uploads\/sites\/74\/2020\/03\/Set4.jpg\" target=\"_blank\" rel=\"noopener noreferrer\">4-UV<\/a><\/td>\n<td style=\"width: 14.2857%\"><a href=\"https:\/\/wordpress-projects.wolfware.ncsu.edu\/bio-183-lab-yhnn8gz\/wp-content\/uploads\/sites\/74\/2020\/03\/Set5.jpg\" target=\"_blank\" rel=\"noopener noreferrer\">5-UV<\/a><\/td>\n<td style=\"width: 14.2857%\"><a href=\"https:\/\/wordpress-projects.wolfware.ncsu.edu\/bio-183-lab-yhnn8gz\/wp-content\/uploads\/sites\/74\/2020\/03\/Set6.jpg\" target=\"_blank\" rel=\"noopener noreferrer\">6-UV<\/a><\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<p>&nbsp;<\/p>\n<h3><span style=\"color: #000000;font-family: UniversCondensed, Arial, sans-serif;font-size: 21px;font-weight: bold\">Calculate the Transformation efficiency using the data from the +LB\/amp\/ara agar plate.<\/span><\/h3>\n<ul>\n<li><span style=\"font-size: 12pt\">Follow the directions one pages 35-37 to calculate your transformation\u00a0 efficiency.\u00a0 Be sure to include the units as you set up each equation.\u00a0 \u00a0You will not be able to fill in the Class Results (Table 2).<\/span><\/li>\n<li><span style=\"font-size: 12pt\">Answer the questions on page 39.\u00a0 You will not be able to answer question 2, but do answer #3 in relation to the ideal transformation efficiency (#1) found on page 35.<\/span><\/li>\n<\/ul>\n<p>&nbsp;<\/p>\n<h3 class=\"style10\"><a href=\"http:\/\/www.youtube.com\/watch?v=4PKjF7OumYo&amp;feature=related\">Molecular Visualization of DNA\u00a0<\/a><\/h3>\n<p class=\"style10\"><span style=\"color: #000000;font-size: 12pt\">0-1:48 = DNA Structure &#8212; Also seen in the Mitosis and Meiosis lab<\/span><\/p>\n<p class=\"style10\"><span style=\"color: #000000;font-size: 12pt\">1:48-2:53 = DNA Replication &#8211;Can you relate this to DNA synthesis? = S phase that you leaned about in the Mitosis and Meiosis in lecture<\/span><\/p>\n<p class=\"style10\"><span style=\"color: #000000;font-size: 12pt\">2:53-4:48 = Transcription &#8211; (you will learn about this and Translation later in lecture)<\/span><\/p>\n<p class=\"style10\"><span style=\"font-size: 12pt\"><span style=\"color: #000000\">4:48-6:57 = Translation &#8211; <\/span><span style=\"color: #000000\">(you will learn about this and Translation later in lecture)<\/span><\/span><\/p>\n<p class=\"style10\"><span style=\"color: #000000;font-size: 12pt\">6:58-7:47 = Hemoglobin and Sickle Cell Anemia\u00a0 -not narrated<\/span><\/p>\n<p>[callout heading=&#8221;DNA Presentations will be compiled by Table Groups in PowerPoint and recorded with both audio and video via Zoom. Share the Zoom video with your TA due by Apr. 13-17.&#8221; headingicon=&#8221;noicon&#8221; textalign=&#8221;textcenter&#8221; type=&#8221;basic&#8221; bgcolor=&#8221;huntyellow&#8221;]To practice your presentations skills, each group of students will receive a DNA Topic and Target Audience to present to the class.  These presentations should run 5-10 minutes.  Each individual will need to turn in a self assessment of their own presentation skills.  See the links below for more information.   This will help you better organize and present your Final Bacterial Project to the class.[\/callout]<\/p>\n<h3 class=\"style9\"><a href=\"https:\/\/wordpress-projects.wolfware.ncsu.edu\/bio-183-lab-yhnn8gz\/wp-content\/uploads\/sites\/74\/2020\/03\/BIO-183-DNA-Presand-Self-Assess-Sp2020-1.pdf\" target=\"_blank\" rel=\"noopener noreferrer\">DNA Presentation Expectations and Self Assessment<\/a> &#8211; 30 points<\/h3>\n<h3 class=\"style9\"><a href=\"https:\/\/wordpress-projects.wolfware.ncsu.edu\/bio-183-lab-yhnn8gz\/wp-content\/uploads\/sites\/74\/2020\/04\/DNAPresentationSheetTA_Sp2020.docx\" target=\"_blank\" rel=\"noopener noreferrer\">DNA Presentation Evaluation Sheet<\/a><\/h3>\n<p>&nbsp;<\/p>\n<h3><span style=\"color: #000000\">Link to information on a Minor in the<\/span> <a href=\"http:\/\/biotech.ncsu.edu\/minors-and-certificate\">Biotechnology Program at NCSU<\/a>\u00a0<span style=\"color: #000000\">and<\/span>\u00a0<a href=\"http:\/\/biotech.ncsu.edu\/courses\">Courses in Biotechnology (BIT) at NCSU<\/a><\/h3>\n<h3 class=\"style11\"><span style=\"color: #000000\">Link to the<\/span>\u00a0<a href=\"http:\/\/www.btec.ncsu.edu\/\">BTEC center on Centennial Campus NCSU\u00a0<\/a><span style=\"color: #000000\">&#8211; where industry and education meet.<\/span><\/h3>\n<h3 class=\"style11\"><span style=\"color: #000000\">Check out the opportunities for undergraduate students! Certificate, Minor, Courses&#8230;..(look under Academic Programs)<\/span><\/h3>\n<h3 class=\"style11\"><span style=\"color: #000000\">At BTEC they use similar transformation procedures to engineer bacteria and yeasts to make (biomanufacturing) chemicals, medicines, vaccines, enzymes, proteins, biofuels&#8230;&#8230;.<\/span><\/h3>\n<h3 class=\"style11\"><span style=\"color: #000000\">Great Short articles related to this important technology !<\/span><\/h3>\n<p class=\"style11\"><a href=\"http:\/\/www.genomecompiler.com\/beautiful-biotechnology-color-changing-flowers\/\">Color-Changing Flowers &#8211; Biotechnology for Everyone<\/a> &#8211; A good short discussion GMOs, mechanics on what was done to change color in Petunias &#8211; the first genetically modified flower commercially available.<\/p>\n<p class=\"style11\"><a href=\"http:\/\/blogs.smithsonianmag.com\/science\/2013\/09\/could-panda-poop-be-the-secret-to-more-efficient-biofuel\/?utm_source=smithsoniantopic&amp;utm_medium=email&amp;utm_campaign=20130915-Weekender\">Could Panda Poop Be the Secret to More Efficient Biofuels?<\/a>\u00a0&#8211; Short article about using genes from bacteria found in Panda feces and inserting them into yeast to increase biofuel efficiency.<\/p>\n","protected":false},"excerpt":{"rendered":"<p>Gene Transformation (2 weeks of lab) &nbsp; \u00a0 &nbsp; Week 1 Background: How GFP is used in research\u00a0(10:03) Funding Basic Science to Revolutionize Medicine\u00a0(3:59) that show how the focus in research has changed over the past 50 years. Stanford Scientists Produce Cancer Drugs from Rare Plants\u00a0(1:59) by making plasmids and inserting them into more easily [&hellip;]<\/p>\n","protected":false},"author":9,"featured_media":0,"parent":0,"menu_order":15,"comment_status":"closed","ping_status":"closed","template":"page-templates\/page-full-width.php","meta":{"_acf_changed":false,"footnotes":""},"class_list":["post-115","page","type-page","status-publish","hentry"],"acf":[],"_links":{"self":[{"href":"https:\/\/wordpress-projects.wolfware.ncsu.edu\/bio-183-lab-yhnn8gz\/wp-json\/wp\/v2\/pages\/115","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/wordpress-projects.wolfware.ncsu.edu\/bio-183-lab-yhnn8gz\/wp-json\/wp\/v2\/pages"}],"about":[{"href":"https:\/\/wordpress-projects.wolfware.ncsu.edu\/bio-183-lab-yhnn8gz\/wp-json\/wp\/v2\/types\/page"}],"author":[{"embeddable":true,"href":"https:\/\/wordpress-projects.wolfware.ncsu.edu\/bio-183-lab-yhnn8gz\/wp-json\/wp\/v2\/users\/9"}],"replies":[{"embeddable":true,"href":"https:\/\/wordpress-projects.wolfware.ncsu.edu\/bio-183-lab-yhnn8gz\/wp-json\/wp\/v2\/comments?post=115"}],"version-history":[{"count":83,"href":"https:\/\/wordpress-projects.wolfware.ncsu.edu\/bio-183-lab-yhnn8gz\/wp-json\/wp\/v2\/pages\/115\/revisions"}],"predecessor-version":[{"id":1279,"href":"https:\/\/wordpress-projects.wolfware.ncsu.edu\/bio-183-lab-yhnn8gz\/wp-json\/wp\/v2\/pages\/115\/revisions\/1279"}],"wp:attachment":[{"href":"https:\/\/wordpress-projects.wolfware.ncsu.edu\/bio-183-lab-yhnn8gz\/wp-json\/wp\/v2\/media?parent=115"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}