Transcript AP-ppt-PCR
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? 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 Sometimes there just isn’t enough Of the DNA we want to work with! The answer is PCR Polymerase Chain Reaction http://dnalc.bii.a-star.edu.sg/shockwave/pcranwhole.htm l DNA sequencing http://www.pbs.org/wgbh/n ova/genome/sequ_flash.ht ml Southern Blotting-used to identify genes/sequences http://dnalc.bii.a-star.edu.sg/shockwave/southan.htm Copy DNA(cDNA) 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 tissues Healthy vs diseased tissues RFLP’s-Restriction Fragment Length Polymorphisms Variations in the length of fragments resulting from action by a specific restriction enzyme uses transposons “Jumping genes” Approx. 45% of human genome Can cause mutations Can be used to trace genetic origin 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 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”?