1. DNA Extraction http://www.vivo.colostate.edu/hbooks/genetics/biotech/manip/conc.html

2. Definition DNA Extraction is the isolation and purification of DNA (deoxyribonucleic acid) http://academy.d20.co.edu/kadets/ lundberg/images/dna.gif

3. Examples DNA extraction is used to isolate…  Mitochondrial DNA  Genomic DNA DNA can be extracted from…  Cells or tissues  Environmental samples http://www.davidkfaux.org/shetlandislandsmtDNA.html http://faculty.uca.edu/~benw/biol1400/pictures/

4. Non-examples DNA Extraction is not used to…  Isolate proteins or RNA  Give information about gene expression http://iwrwww1.fzk.de/biostruct/

5. Downstream Applications After DNA is extracted, it is used as a template in further molecular techniques such as…  PCR (polymerase chain reaction)  RFLP (restriction fragment length polymorphism)  Southern Blotting

6. DNA Analysis Downstream techniques can:  Reveal how organisms are related  Identify cryptic species  Locate mutations in DNA http://www.biocompare.com/itemdetails.asp?itemid=687320 http://www.cdc.gov/hemochromatosis/training/images/dna.jpg

7. Quick Quiz What is the purpose of DNA extraction? a. To learn how to safely work with chemicals b. To quantify the amount of DNA per cell c. To determine how organisms are related d. To purify DNA from cells

8. Qiagen DNA Extraction Qiagen DNA Extraction Tissue Lysis. Detergents lyse membranes. Proteinase K breaks down proteins. Sample loading onto column. Nucleic acids bind to membrane and other components flow through. Column washing. Removes any contaminants. Elution of DNA. DNA is released from column and ready for use in downstream applications.

9. Quick Quiz The purpose of the column is to: a. Separate DNA from RNA b. Separate DNA molecules from cellular debris c. Remove bacterial contaminants d. Help suspend DNA in elution buffer

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11. Resources DNA Extraction Handout DNEasy Tissue Kit Handbook http://www1.qiagen.com/literature/protocols/ DNeasyTissue.aspx

12. California State Chemistry Standards Grade 8 6c. Living organisms have many different kinds of molecules… Grades 9-12 6. Solutions are homogenous mixtures of two or more substances 7. Energy is exchanged or transformed in all chemical reactions and physical changes of matter 8. Chemical reaction rates depend on factors that influence the frequency of collision of reactant molecules

13. California State Biology Standards Grade 7 2e. DNA is the genetic material of living organisms and is located in the chromosomes of each cell 3a. Genetic variation and environmental factors are causes of evolution and diversity of organisms Grades 9-12 2. Mutation and sexual reproduction lead to genetic variation in a population 7. The frequency of an allele in a gene pool of a population depends on many factors and may be stable or unstable over time

14. California State Investigation and Experimentation Standards Grades 7, 9-12 a/b. Select and use appropriate tools and technology to perform tests, collect data, analyze relationships, and display data Grade 8 a. Plan and conduct a scientific investigation to test a hypothesis

15. National Standards Grades 6-12 Content Standard A: Science as Inquiry Content Standard C: Life Science Content Standard E: Science and Technology