BIOL 5060- Midterm Exam Guide - Comprehensive Notes for the exam ( 171 pages long!)
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Common mistakes with pcr: oligo should be ~18-24, optimal temp. Looking at the structural analysis of dna: purpose= allows us to examine a specific region of dna in a large population of sequences, wild-type genome vs. deletion how do we know which we have, 1. Southern blots- looking for something where we don"t know what to expect- way of examining the dna: method: 1. isolate large mw dna 2. Cut with a restriction enzyme (complete digest with something that will cut less frequently) 3. Run digest on a gel (after these 3 steps we"ve separated dna based on the size of the fragments collectively separates various regions of the genome into discrete fragments) 4. Transfer dna to a filter/ membrane : denature the dna (so its not double stranded and amenable for base pairing) 6. Probe with a labeled, denatured dna molecule 7. Wash probe to remove non- specifically bound material 8.
