MT2practicef11 - MCB160L- EXAM 2 Practice 2011 The topics...

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MCB160L- EXAM 2 Practice 2011 The topics for the exam listed below are the same as in the separate posted file. The practice exam follows. The majority of the topics for Exam 2 were covered in Lectures 6 – 12; however in this exam you will also be tested on the concepts from Lectures 4-5 that were not covered in Exam 1, for example blue- white screening, directional cloning and possible ligation products (see list). Also keep in mind that some material from Exam 1 serves as building blocks for later work- so if you struggled with basic concepts on that exam (e.g. fly genetics, basic cloning) you should review that material as well. Material from the DataRec/Worksheets and manual corresponding to the lectures/topics covered may also be included on the exam (Worksheet M4, Worksheet M5 ,DataRecord M5, Data Record/Worksheet D2, Computer exercise M4, Cloning Report part 1, Worksheet A1, Drosophila Report 1. This topic list includes the main topic areas that we covered but does not necessarily include all aspects of these areas (other subtopics within these lectures and worksheets should also be studied). Yeast and C. elegans are NOT included on the exam. A practice exam from a previous quarter has been posted that covers similar material. It will give you an idea of the types of questions that could be asked, but of course cannot cover all of the topics that may be tested. (A.) Molecular Biology Subcloning-there will be some overlap with the first exam in this category (e.g. ligation) but overall Exam 2 will concentrate on the application of the concepts to cloning experiments. Ligation theory and practice: Parameters that affect the outcome of ligation reactions -how to minimize recircularization, phosphatase, directional cloning vs. non-directional cloning. Possible products in ligation reactions and conditions that favor recovery of one product vs another Problems resulting from partial digests Directional cloning and its applications, including fusion proteins Rationale for blue/white screening for inserts Gel purification of DNA fragments, advantages of this approach for obtaining desired products in subcloning strategies Minipreps- rationale for steps during plasmid DNA isolation procedure PCR- Reaction components and general steps in the reaction cycle (denaturation, primer annealing, primer extension) PCR mechanism during first 3 cycles (primer position and polarity, types of products, etc) Primer structure and design considerations, contamination issues Human DNA genotyping by VNTR analysis B. Drosophila Drosophila Exp. 1.1 and 1.2 Exceptional vs mendelian progeny from cross 1.1 Rationale for choosing males to use in cross 1.2 Possible genotypes of cross 1.2 progeny for pw+ located on different chromosomes Enhancer traps, GAL4-UAS trans-activation system in Dros Exp 1 and applications Effect of P-element mobilization on germline and somatic cells and possible outcomes (eg. how
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MT2practicef11 - MCB160L- EXAM 2 Practice 2011 The topics...

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