1 / 22

Cloning

Last class. Cloning. Transformation. Paper 2 intro. Plasmid prep. Plasmid in cells. How to get out?. Plasmid prep. Plasmid prep: 1 Lyse the cell. Plasmid prep: 2 Precipitate Proteins. Plasmid prep: 3 Bind plasmid. Plasmid prep: 3 Bind plasmid. Plasmid prep: 3 Bind plasmid.

halima
Download Presentation

Cloning

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. Last class Cloning Transformation Paper 2 intro

  2. Plasmid prep Plasmid in cells How to get out?

  3. Plasmid prep

  4. Plasmid prep: 1 Lyse the cell

  5. Plasmid prep: 2 Precipitate Proteins

  6. Plasmid prep: 3 Bind plasmid

  7. Plasmid prep: 3 Bind plasmid

  8. Plasmid prep: 3 Bind plasmid

  9. Plasmid prep: 4 Elute plasmid

  10. Plasmid prep & RE mapping Lyse bacteria, isolate plasmid pBK-CMV with gene RE digest and run on a gel Determine RE map of plasmid

  11. First experiments Purpose? Experiment? Results?

  12. First experiments Purpose? Experiment? Results? Next step?

  13. c-Myc? Purpose? Experiment? Results?

  14. Control: Prediction for iPSC expression pattern Purpose? Experiment? Prediction for miR iPSCs?

  15. Control: Prediction for exogenous genes? Purpose? Experiment? Prediction for miR iPSCs? What else?

  16. miRNA iPSCs? Purpose? Experiment? Prediction for miR iPSCs?

  17. ChIPs? Purpose? Experiment?

  18. ChIP Y Y Y Y Y Y Unbind & sequence

  19. ChIPs? Purpose? Experiment? Result?

  20. ChiPs + Expression = ?

  21. % GFP +ve colonies = ? Purpose? Experiment? Results?

  22. Issues? Controls? Experiments?

More Related