User:Karmella Haynes/Notebook/ASU courses/2012/01/25

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1/25/12

  • Minipreps: O2-GFP, O1-GFP, Osym-GFP, LacI-O2 for minipreps (5 mL)
  • Transformations: co-transform reporter/ LacI



Minipreps
> Measure DNA concentration using the Take3 plate (Sarah's Odyssey system)

Sample OD 260 260/280 ng/μL
1. O2-GFP 1.613 2.208 80.665
2. O1-GFP 1.911 2.151 95.538
3. Osym-GFP 1.019 2.154 50.961
4. LacI-O2 1.189 2.214 59.459



Transformation
> Use 25 ng of each plasmid

Plasmid 1 μL Plasmid 2 μL
1. O2-GFP 0.3 LacI-O2 0.4
2. O1-GFP 0.3 LacI-O2 0.4
3. Osym-GFP 0.5 LacI-O2 0.4

> Add 50 μL NEB10B cells (quick and dirty transformation); plate on 100 μg/mL Amp + 50 μg/mL Kan LB agar


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