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Review Sheet for Final Exam --- New Material Bio310 S’09 Chapter 16 Why are bacteria often used to engineer new DNA molecules or to express useful proteins? What is a restriction enzyme? Where did we borrow them from? In working with molecular biology, what is meant by a probe? A primer? What is a Southern blot? What is meant by plasmid cloning? How does PCR work? Be able to describe the steps you would go through to clone a eukaryotic gene into a bacterium and find a correct copy of the plasmid carrying it. How does whole genome shot-gun sequencing work? What is meant by annotation in reference to genomics? How does a gene chip measure transcriptional levels in a cell? Chapter 17 (bits of 18 &19 ) Differentiate between distance matrix and maximum parsimony methods in building a phylogeny. How does one use 16 S rRNA for phylogenetics? How does horizontal gene transfer complicate phylogenetics for bacteria and how do you get around this problem? Know the different ways to identify unknown bacteria. Compare and contrast archaea and bacteria What is meant by an extremophile? Why are we worried that archaea may help speed up global warming? Are most archaea extremophiles? Where would you expect to find a lot of proteobacteria? What is a serovar? What is an indicator bacterium? Chapter 24 How does FISH work? What does it help you do? What is the difference between direct counts and viable counts? How can you make direct counts more accurate for quantifying living cells? Why can’t we culture most bacteria in a lab? What is meant by the term “microbial succession” and why is it important to human health? What is a biofilm? Why are they important? Chapter 26 What do we mean by an “efficient parasite”? Be able to give an example on both sides. What is meant by the concept of “probiotic”? Know the types of innate defenses. What is the purpose of the inflammation response? How does phagocytosis work? How do some pathogens get around it? Be able to distinguish between the classes of pathogens How do pathogens increase their virulence? What is the difference between an endotoxin and an exotoxin? Lab topics (Experiments 8-10) In our conjugation experiment, what two things had to happen to see an erythromycin resistant colony of Flavobacterium appear on the selective plates (PYE-2+ Erm)? Most of the Flavobacterium colonies on the PYE-2 plates were motile, but a few groups had one or two non-motile colonies. What made those colonies non-motile and why was it significant? Why didn’t the LB plates used in the transformation just turn out confluent? How, basically, does the miniprep work? (just essential details, no amounts of reagents needed) What is the purpose of the RNase in the miniprep protocol? If you counted 225 plaques in your 1/100,000 dilution plate, how many PFU/ml in the original sample?