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Last class Cloning Transformation Paper 2 intro.

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Presentation on theme: "Last class Cloning Transformation Paper 2 intro."— 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: 2 Precipitate Proteins & DNA

5 Plasmid prep: 3 Bind plasmid

6 Plasmid prep: 3 Bind plasmid

7 Plasmid prep: 3 Bind plasmid

8 Plasmid prep: 4 Elute plasmid

9 Plasmid prep & RE mapping
Lyse bacteria, isolate plasmid pBK-CMV with gene RE digest and run on a gel Determine RE map of plasmid (size of insert = 5.5Kb)

10 Implied knowledge! What were some things you had to look up?

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 ChIP Y Y Unbind & sequence

18 ChIPs? Purpose? Experiment? Result?

19 ChiPs + Expression = ?

20 % GFP +ve colonies = ? Purpose? Experiment? Results?

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