1. Biotechnology Copyright © Texas Education Agency 2009. All rights reserved.

2. How do they do that!? zHow do crime scene investigators use such small samples of blood and tissues to solve crimes? zHow do scientists make mice glow? Lets find out…

3. CRIME SCENE 1.Collect tissue samples. 2.Extract and Purify DNA 3.Amplify (make lots of copies of your sample) using PCR. 4.Cut DNA with restriction enzymes 5.Run gel electrophoresis to make a DNA fingerprint 6.Compare fingerprints to identify suspects

4. CRIME SCENE 1.Collect tissue samples. What kinds of samples can be used to extract DNA?

5. Extract & Purify your DNA zJust like you did in your DNA extraction LAB!!!

6. Polymerase Chain Reaction zMethod used to make lots of copies of a section of DNA zAMPLIFICATION

7. PCR zhttp://www.dnalc.org/resources/animatio ns/pcr.htmlhttp://www.dnalc.org/resources/animatio ns/pcr.html

8. Restriction Enzymes Copyright © Texas Education Agency 2009. All rights reserved. Restriction Enzyme Animation

9. zRestriction enzyme – an enzyme which binds to DNA at a specific base sequence and then cuts the DNA Restriction Enzymes Copyright © Texas Education Agency 2009. All rights reserved.

10. zAfter cutting DNA with restriction enzymes, the fragments can be separated on an agarose gel. zThe smaller fragments will migrate further than the longer fragments in an electric field. Restriction Enzymes Copyright © Texas Education Agency 2009. All rights reserved.

11. Agarose Gel Electrophoresis zElectrophoresis is a molecular technique that separates nucleic acids and proteins based on: Size and +-+ Charge +-+ Copyright © Texas Education Agency 2009. All rights reserved.

12. zDNA is a negatively charged molecule and therefore is attracted to positive charges. Agarose Gel Electrophoresis Copyright © Texas Education Agency 2009. All rights reserved.

13. zAgarose provides a matrix through which DNA molecules migrate.  Larger molecules move through the matrix slower than small molecules Agarose Gel Electrophoresis Copyright © Texas Education Agency 2009. All rights reserved.

14. Agarose Gel Electrophoresis Copyright © Texas Education Agency 2009. All rights reserved. http://www.dnalc.org/resources/ani mations/gelelectrophoresis.html http://www.sumanasinc.com/ webcontent/animations/conten t/gelelectrophoresis.html

16. RFLP’s zhttp://highered.mcgraw- hill.com/classware/ala.do?isbn=00729658 19&alaid=ala_1032447&showSelfStudyTre e=truehttp://highered.mcgraw- hill.com/classware/ala.do?isbn=00729658 19&alaid=ala_1032447&showSelfStudyTre e=true

17. Applications of Gel Electrophoresis zIdentifying genes for study or transfer zSeparating DNA products zDNA fingerprinting yCrime scene analysis yPaternity

18. Who did it?

19. Who's the Daddy?

20. Recombinant DNA zRecombinant DNA technology  Allows DNA to be combined from different sources  Also called genetic engineering or transgenics Copyright © Texas Education Agency 2009. All rights reserved.

21. zVector – DNA source which can replicate and is used to carry foreign genes or DNA fragments. zRecombinant DNA – A vector that has taken up a foreign piece of DNA. Recombinant DNA Copyright © Texas Education Agency 2009. All rights reserved.

22. zRecognition site – specific base sequence on DNA where a restriction enzyme binds. Restriction Enzymes Copyright © Texas Education Agency 2009. All rights reserved.

23. zMost restriction enzymes cut within the recognition site. zWhen restriction enzymes cut in a zig zag pattern, sticky ends are generated. Copyright © Texas Education Agency 2009. All rights reserved. Restriction Enzymes

24. zOverhanging sticky ends will complementarily base pair, creating a recombinant DNA molecule. zDNA ligase will seal the nick in the phosphodiester backbone. Restriction Enzymes Copyright © Texas Education Agency 2009. All rights reserved.

25. Transformation zTransformation – the process by which organisms take up and express foreign DNA Copyright © Texas Education Agency 2009. All rights reserved.

26. zBacterial Transformation  Bacteria, such as E.coli, can take up and express foreign DNA, usually in the form of a plasmid. Transformation Copyright © Texas Education Agency 2009. All rights reserved.

27. zGene cloning – using bacterial transformation to make lots of copies of a desired gene. Transformation Copyright © Texas Education Agency 2009. All rights reserved. Gene Cloning Animation

28. Producing insulin with bacteria