1. DNA& Biotechnology

2. I. What is DNA? Deoxyribonucleic acid Polymer Made of long chains of nucleotides – Phosphate group, sugar, & a nitrogen base 4 bases: C, G, A, T – C  G, G  C – A  T, T  A – Bases held together by hydrogen bonds 1

3. What is the structure of DNA? 2 strands running opposite each other – Sides are alternating deoxyribose sugar & a phosphate molecule - N-bases are connected to the sugar Form a twisting ladder, double helix 23

4. DNA’s job DNA is transcribed into RNA which codes for amino acids and makes proteins Not all DNA codes for proteins Coding sequences = exons Noncoding parts = introns RNA splicing trims out introns 4 p. 14-16 of NG, 30-1

5. Where is DNA found? Coiled up –making chromosomes – Found inside the nucleus of cells 5 p.6-7 NG

6. Biotechnology The study & manipulation of living things or their component molecules, cells, etc. biologycorner.com

7. Early biotech Planting crops Breeding animals to get specific traits Using yeast to turn – Fruit juice into wine – Malt and hops into beer – Making bread rise Using bacteria to turn milk into yogurt and cheese From Amanda Noller via Pamela Peters, from Biotechnology: A Guide To Genetic Engineering. Wm. C. Brown Publishers, Inc., 1993.

8. Current uses of biotech Amplifying DNA to see it Sequencing genomes Gene splicing Recombinant DNA/ Transformation Creating effective medicines Genetic Engineering Cloning Stem Cell Research And so much more… http://www.iptv.org/exploremore/ge/what/insulin.cfm From Amanda Noller via Pamela Peters, from Biotechnology: A Guide To Genetic Engineering. Wm. C. Brown Publishers, Inc., 1993

9. Isolating & manipulating DNA Why? Changing DNA nucleotides can alter proteins – Making medicines/hormones ex: insulin p.64 – http://www.youtube.com/watch?v=tJP_6nAPES4 http://www.youtube.com/watch?v=tJP_6nAPES4 chemicalconnections.org

10. Chromatography a set of techniques used to separate different compounds. Uses: isolating new compounds, analyzing subtle differences between different samples, and even in the sequencing of DNA.

11. Paper chromatography stationary phase (a solid, or a liquid supported on a solid) mobile phase (a liquid or a gas). The mobile phase flows through the stationary phase and carries the components of the mixture with it. Different components travel at different rates. In paper chromatography, the stationary phase is a uniform absorbent paper. The mobile phase is a suitable liquid solvent or mixture of solvents.

12. R f values Some compounds in a mixture travel almost as far as the solvent does; some stay much closer to the base line. The distance travelled relative to the solvent is called the R f value. For each compound it can be worked out using the formula: R f = distance traveled by compound distance traveled by solvent For example, if one component of a mixture travelled 9.6 cm from the base line while the solvent had travelled 12.0 cm, then the R f value for that component is: 9.6/12 = 0.8

13. Gel electrophoresis Separation technique Visual way to see the pieces of DNA Uses electrical impulses to pull the DNA through a matrix like jello (called Agarose) DNA in negatively charged, so it moves towards a positive pole and away from the negative Separated by size, shape, & charge http://www.dnalc.org/resources/animations/gelelectrophoresis.html

14. The cut DNA is mixed with blue dye and loaded into a gel 6

15. DNA moves towards positive end (due to the negative charges of the phosphates) The dye moves faster than the smallest DNA fragments Smaller the fragments the faster they move 7

16. Using a micropipetter http://www.youtube.com/watch?v=uEy_NGDfo_8 Set desired volume by turning the dial Max volume is on top of pipette Put sterile tip on from box To load sample, press down until 1 st stop Release button to draw up sample To release sample into container, push down on button, past the 1 st stop Withdraw pipette from container before releasing the button

17. PCR Polymerase chain reaction Used for forensics, medicine, research, and more Amplifies small sections of DNA to get LOTS of DNA

18. PC R 8

19. What you need DNA - you need something to copy! Taq polymerase - copies the DNA Primers - get the polymerase going dNTP - the As, Ts, Cs, and Gs buffered (pH controlled) tube of water thermocycler

20. Taq polymerase 10 11 12 13

21. In the thermocycler… 1.Heat to 95ºC (Denature) DNA denatures into its two strands 2.Cool down to 58ºC- 65°C (Annealing) Primer sequences stick to the template 3.Heat to 72ºC (Synthesis) Taq polymerase attach at the primer and make a complimentary copy of the DNA 4.Repeat

22. Start with one copy – One copy, one cycle- 2 copies – 2 copies, one cycle- 4 copies – 4 copies, one cycle- 8 copies Typical runs include between 25-35 cycles Finish with billions of identical copies of the original DNA fragment http://learn.genetics.utah.edu/content/labs/pcr/

23. Sources for pictures 1. http://www.scq.ubc.ca/a-monks-flourishing-garden-the-basics-of-molecular- biology-explained/ 2. 3. http://biolibogy.com/chemistryoflife.htmlhttp://biolibogy.com/chemistryoflife.html 4.http://publications.nigms.nih.gov/thenewgenetics/chapter1.html 5.http://publications.nigms.nih.gov/thenewgenetics/chapter1.html#dnastr 6-9. Amanda Noller’s Biotechnology ppt 12. http://andrew.cmu.eduhttp://andrew.cmu.edu 13. http://coloradoindependent.comhttp://coloradoindependent.com