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

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

Wednesday Thursday 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

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!!!

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

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

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

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