StudyGuide MidF09

StudyGuide MidF09 - STUDY GUIDE Molecular Biology Midterm...

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STUDY GUIDE - Molecular Biology Midterm Fall 2009 MIDTERM EXAM is Oct 5-7 in class. Review session: Sunday Oct 4 at 4 PM in the lab. If it is too crowded we will move and I'll write the room number on the board. Please come with QUESTIONS (don’t expect me to repeat the first half of the semester). If you cannot make the review and you have questions, let me know and we will try to set up a time to meet. This exam will cover both theory and practice of the techniques we have performed this semester. You will not have to demonstrate techniques such as loading a gel. It will not include anything on 16s rDNA. M, T, W sections have all covered the same topics, but perhaps not in the same order or depth, so it is important that each student understand what we covered. Lab #1, safety handout: Proper use and care of the micropipettors. Choosing the correct micropipettor. Determining the volume setting. How to draw up a sample. How to properly use the microfuge and other lab equipment. Understand how to work safely in the lab and how to handle problems such as broken glass or an alcohol fire. Use of bacteria in the Molecular Biology lab: How to streak plates (with a plastic loop!). Understand how antibiotic resistance is achieved. Be able to explain our plating results with antibiotic resistant bacteria. Aseptic techniques and safe handling (importance of sterilized broths, agar, loops, pipette tips etc). Proper disposal of recombinant bacteria. Importance of using bacteria non-pathogenic to humans. Why are bacteria the workhorses of a modern molecular biology lab? Bacterial culture techniques ( E. coli growth curve): What growth phase is best for making competent cells or for isolating plasmid DNA? What techniques use broth cultures vs. individual colonies? When do we streak a plate and when do we spread? Lab #2, cloning experiment: Understand how a gene can be cloned into a plasmid vector. Understand our cloning scheme. What bands to expect from restriction enzyme digestion of the starting plasmid pGEM and the successful pIg recombinant. Understand all steps
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StudyGuide MidF09 - STUDY GUIDE Molecular Biology Midterm...

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