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Lecture

Lec21_140303.pdf

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Department
Statistics
Course
STAT151
Professor
Gregory Wagner
Semester
Winter

Description
BIOL207 Lec21 2014-03-03 CHAPTER 8 A. Molecular Biology a. study macromolecules (DNA, RNA, proteins, etc.) physically and directly, rather than making inferences based on patterns of inheritance B. DNA extraction a. lyse tissue in extraction buffer containing minimum: i. detergent (e.g. SDS) ii. salts (e.g. NaCl) iii. chelating agent (e.g. EDTA) b. precipitate proteins and other debris from lysate; retain supernatant c. precipitate DNA from supernatant using alcohol (e.g. ethanol); discard supernatant d. dissolve (“resuspend”) DNA pellet in pure water i. maybe with some: salt, chelating agent, pH buffer C. PCR a. used both analytically (detect DNA with specific sequence) and preparatively (make more DNA with specific sequence) b. in vitro DNA replication c. specificity comes from primers; all DNApol need a primer d. PCR reaction components i. template DNA (usually genomic DNA) ii. pair of primers iii. nucleotides (dNTPs) iv. buffer (salts, Mg , pH buffer) v. DNA polymerase (usually thermostable, e.g. Taq) e. reaction goes in a thermal cycler for successive rounds of i. denaturation, annealing, extension http://www.youtube.com/watch?v=2KoLnIwoZKU http://www.le.ac.uk/richardiii/science/extractionofdna.html D. Restriction Digestion http://www.youtube.com/watch?v=
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