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Tutorial_2___Isolation_of_Genomic_DNA - BIOL 208 ...

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Unformatted text preview: BIOL 208 Tutorial 2 – Isolation of Genomic DNA Before Tutorial: Gather Data ⋅ Go to FlyBase or WormBase and familiarize yourself with the content of the databases. Click around and explore! During Tutorial: Data Analysis 1. What types of molecular analyses require purified DNA? 2. You want to begin cloning your gene of interest. It is always a good idea to start out with more DNA than you think you will need for one ligation. What are some of the reasons for this? List at least 5 reasons. You estimate that you need 250 μg of DNA for cloning. a. How much extra DNA should you prepare? Discuss these factors with your group. b. Find a protocol for genomic DNA isolation on FlyBase or WormBase (depending on which organism your gene originates from). Estimate how many flies or worms you would need to use to collect enough genomic DNA for your experiments? Find a citation for this information. c. How do you maintain this many worms/flies? Describe the time involved and space/food requirements. Thinking Question: Due the following Monday by Noon ⋅ Molecular biologists use the term “purify” a lot (ie: I purified my protein!). What does it mean to purify genomic DNA? ⋅ To evaluate the quality of the purified uncut genomic DNA on an agarose gel: ⋅ What concentration of agarose would you use? (high or low) Why? ⋅ If the purification had gone well, what would you see and why? ⋅ If nucleases had contaminated the extraction buffer, what would you see and why? ...
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