2. Problems Not all bacteria pick up plasmid-how do we distinguish? Annealing of human DNA to plasmid is random-how do we distinguish which plasmids have human DNA?

4. How do we find the gene we want in a library? Probing! It’s like fishing For DNA Relies on complementarity of The double strand

5. http://dnalc.bii.a-star.edu.sg/shockwave/pcranwhole.htm l Sometimes there just isn’t enough Of the DNA we want to work with! The answer is PCR Polymerase Chain Reaction

6. http://www.pbs.org/wgbh/n ova/genome/sequ_flash.ht ml http://www.pbs.org/wgbh/n ova/genome/sequ_flash.ht mlDNAsequencing

7. http://dnalc.bii.a-star.edu.sg/shockwave/southan.htm Southern Blotting-used to identify genes/sequences

8. Copy DNA- (cDNA)

9. DNA microarray analysis Microscopic DNA attached to solid surface like glass, silicon Allows us to answer questions about gene activity Different stages of development Different stages of development Different tissues Different tissues Healthy vs diseased tissues Healthy vs diseased tissues

10. RFLP’s-Restriction Fragment Length Polymorphisms Variations in the length of fragments resulting from action by a specific restriction enzyme uses

11. transposons “Jumping genes” Approx. 45% of human genome Can cause mutations Can be used to trace genetic origin

12. SAFETY, SAFETY, SAFETY!!! THINK! No food or drink anywhere!!!! Don’t touch face, doorknobs, goggles, books, faucet handles, etc with gloves on Clamshell technique Wipe surfaces thoroughly w/ ethanol spray

13. So what about our lab? We artificially transformed bacteria Gene comes from jelly fish! The Arabinose operon How do we know which bacteria took up plasmid? What is required in the agar to make them “glow”?