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Week 6: Electro-transformation and screening of the genomic library

Week 6: Electro-transformation and screening of the genomic library . Wednesday Look at transformation results Transform the following: Water pUWL500 Library (L1 –L4) Thursday Screen colonies Ex 9A and 10 pt I and II due with the following questions: Ex #9 1,3,4,5 Ex#10  all .

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Week 6: Electro-transformation and screening of the genomic library

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  1. Week 6:Electro-transformation and screening of the genomic library

  2. Wednesday • Look at transformation results • Transform the following: • Water • pUWL500 • Library (L1 –L4) • Thursday • Screen colonies • Ex 9A and 10 pt I and II due with the following questions: • Ex #9 1,3,4,5 Ex#10  all

  3. Wednesday • Transformation of ligation: • We will transform the following, each in 35 uL of competent cells: • 1 uL water • 20ng pUWL500 • 5 uL of the ligation reactions • Remember to dialyze everything before transformation, and plate on LB + Amp!!!

  4. Transformation tips: • Keep cells on ice at all times • Only add cells to cold cuvettes • Immediately add 1 mL SOC media immediately after electroporation • Incubate 30 minutes after adding SOC

  5. Checklist: • Plate the following volumes from the transformations: • Negative control: 100 uL • pUWL500: 100 uL • Ligation reactions: 200 uL for each reaction • Save the ligation transformations for next time • Place plates in 37° incubator

  6. Week 6:Electro-transformation and screening of the genomic library

  7. Last time: • We transformed controls and our ligation reactions. Today we will observe the results.

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