{"id":431618,"date":"2018-10-02T18:14:19","date_gmt":"2018-10-02T18:14:19","guid":{"rendered":"https:\/\/essaypaper.org\/?p=43085"},"modified":"2018-10-24T08:54:22","modified_gmt":"2018-10-24T08:54:22","slug":"subcloning-yeast-two-hybrid","status":"publish","type":"post","link":"https:\/\/www.benedictsol.com\/blogs\/subcloning-yeast-two-hybrid\/","title":{"rendered":"Subcloning &amp; Yeast two-hybrid"},"content":{"rendered":"<div class=\"col-lg-10 col-md-9 col-sm-16\">\n<h2 class=\"form-control-static orderTopic\" title=\"Subcloning &amp; Yeast two-hybrid\">Subcloning &amp; Yeast two-hybrid<\/h2>\n<\/div>\n<div class=\"col-lg-10 col-md-9 col-sm-16\"><label class=\"control-label\">Description<\/label>Title<br \/>\nThis should convey the point of the mini-project in 15 words or less.<br \/>\n**Introduction (about 200 words):<br \/>\nYou can use figures in addition to text in this section if you like.<br \/>\n\u2022 Subcloning<br \/>\n-What are plasmids and how can they be used?<br \/>\n-How can a piece of DNA encoding a protein of interest -be isolated from a<br \/>\nplasmid?<br \/>\n-How can such a DNA fragment be introduced into another plasmid?<br \/>\n\u2022 Yeast two-hybrid<br \/>\n-What techniques can be used to verify that subcloning has successfully<br \/>\nproduced the desired plasmid?<br \/>\n-What are the main applications of this technique?<br \/>\n-How does it work?<br \/>\n\u2022 Finish the Introduction with a very clear statement of the AIM of the Project<br \/>\n**Materials and Methods:<br \/>\nInclude a flow chart of the techniques used<br \/>\nDo NOT write out what you did &#8211; just a summary to demonstrate that you understand the aims<br \/>\nof each prac &amp; how they linked together to form the project.<br \/>\n\uf075 Steps only<br \/>\n\uf075 No details of reagents etc.<br \/>\n**Results (about 300 words plus figures):<br \/>\n-Present and describe your individual results and the compiled class data.<br \/>\n*You will use the notes you made during the practical classes, plus photos of gels and yeast plates, and class transformation\/streaking data.<br \/>\n&#8211; Include in your report figures showing your gels and plates your samples (and what they were) and label the molecular weight markers with the size of each fragment (in nucleotides or kilobases).<br \/>\n*Must be labelled correctly and be described in text.<br \/>\nIn the text of this section, make sure you address the following points:<br \/>\n\u2022 What size fragments did you expect from the restriction digestion of each plasmid in<br \/>\npractical 1? Based on your results, which plasmid did you cut?<br \/>\n\u2022 What PCR and restriction digests patterns did you expect in practicals 2 and 3, if the<br \/>\ncolony you picked contained the desired Bad-pGBT9 plasmid? Do your results match<br \/>\nthese expectations?<br \/>\n\u2022 Which plasmid(s) did you transform into yeast in practical 7? Did any transformant<br \/>\ncolonies grow on the -LW plate? Considering the entire class data, which<br \/>\ntransformations generally yielded colonies on the -LW plates? Explain these results.<br \/>\n\u2022 Which transformants grew on your -LWH plate in practical 9? Did this outcome match<br \/>\nthe whole-class data?<br \/>\n**Conclusion and discussion (about 400 words)<br \/>\n\u2022 Re-state your Aim and describe the extent to which it was achieved.<br \/>\n\u2022 Outline the purpose of including the \u201cnegative control\u201d PCR reaction and culture in<br \/>\npractical 5. Did these behave as expected? If so, what can you conclude the results from the PCR and miniprep from the tubes inoculated with the bacterial colony. If the controls did not yield the expected results, why not?<br \/>\n\u2022 Did the rest of your results match your expectations? If you had any surprising results, suggest possible reasons for these. Was it human error (eg did you not pipette the correct volumes, or threw out the supernatant when you were meant to keep it)? Could defective reagents or equipment be responsible, such as enzymes that had lost activity, PCR machine\/heatblock not achieving specified temperature? (Consider class data in relation to this possibility).<br \/>\n\u2022 Write an overall conclusion describing very briefly what the aim of the project was, what the findings were and how it can be improved in the future.<br \/>\n*******Format*******<br \/>\nat least six different references,<br \/>\nIn-text referencing should occur in the Introduction,<br \/>\nMaterials and Methods and Discussion.<br \/>\nFont: Times New Roman<br \/>\nSize: 12 point<br \/>\nLines: Double spaces<br \/>\nIn-text referencing should occur in the Introduction,<br \/>\nMaterials and Methods and Discussion with the pg no.\n<\/div>\n","protected":false},"excerpt":{"rendered":"<p>Subcloning &amp; Yeast two-hybrid DescriptionTitle This should convey the point of the mini-project in 15 words or less. **Introduction (about 200 words): You can use figures in addition to text in this section if you like. \u2022 Subcloning -What are <a href=\"https:\/\/www.benedictsol.com\/blogs\/subcloning-yeast-two-hybrid\/\" class=\"read-more\">Read More &#8230;<\/a><\/p>\n","protected":false},"author":1,"featured_media":0,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[15],"tags":[],"class_list":["post-431618","post","type-post","status-publish","format-standard","hentry","category-essay-paper-writing"],"_links":{"self":[{"href":"https:\/\/www.benedictsol.com\/blogs\/wp-json\/wp\/v2\/posts\/431618","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/www.benedictsol.com\/blogs\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/www.benedictsol.com\/blogs\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/www.benedictsol.com\/blogs\/wp-json\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"https:\/\/www.benedictsol.com\/blogs\/wp-json\/wp\/v2\/comments?post=431618"}],"version-history":[{"count":0,"href":"https:\/\/www.benedictsol.com\/blogs\/wp-json\/wp\/v2\/posts\/431618\/revisions"}],"wp:attachment":[{"href":"https:\/\/www.benedictsol.com\/blogs\/wp-json\/wp\/v2\/media?parent=431618"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/www.benedictsol.com\/blogs\/wp-json\/wp\/v2\/categories?post=431618"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/www.benedictsol.com\/blogs\/wp-json\/wp\/v2\/tags?post=431618"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}