1 / 13

Problems Not all bacteria pick up plasmid-how do we distinguish? Remember ampR gene?

Problems Not all bacteria pick up plasmid-how do we distinguish? Remember ampR gene? Annealing of human DNA to plasmid is random-how do we distinguish which plasmids have human DNA? Remember location of restriction site Plasmids w/human DNA cannot breakdown lactose, therefore no blue color.

avedis
Télécharger la présentation

Problems Not all bacteria pick up plasmid-how do we distinguish? Remember ampR gene?

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript

  1. Problems • Not all bacteria pick up plasmid-how do we distinguish? • Remember ampR gene? • Annealing of human DNA to plasmid is random-how do we distinguish which plasmids have human DNA? • Remember location of restriction site • Plasmids w/human DNA cannot breakdown lactose, therefore no blue color

  2. So what about our lab? • We are artificially transforming bacteria • Giving them a new gene-it will make them glow in the dark!!! • Gene comes from jelly fish! • Operon includes genes to make GFP AND IS controlled by arabinose, which is an inducer • How do we know which bacteria took up plasmid? • What is required in the agar to make them “glow”?

  3. SAFETY, SAFETY, SAFETY!!! • THINK! • Clamshell technique • Wipe surfaces thoroughly! • No food or drink anywhere!!!! • Don’t touch face, doorknobs, goggles, books, faucet handles, your ass,etc….

  4. How do we find the genewe want in a library?

  5. 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

  6. http://www.pbs.org/wgbh/nova/genome/sequ_flash.html • DNA sequencing

  7. Southern Blotting http://dnalc.bii.a-star.edu.sg/shockwave/southan.htm

  8. Copy DNA- (cDNA) Since bacteria don’t normally Process their mRNA, they have no way of cutting out introns of eukaryotic genes

  9. DNA microarray analysis • Allows us to answer questions about gene activity • Different stages of development • Different tissues • Healthy vs diseased tissues

  10. RFLP’s-Restriction Fragment Length Polymorphismsnon-coding sequences have individual differences

  11. Alu insert

More Related