Transforming Plasmids using Competent cells DH5Alfa

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Guy Sovak
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Transforming Plasmids using Competent cells DH5Alfa

Transforming Plasmids using Competent cells DH5Alfa

Take 50ul of DH5Alfa Competent cells (thaw on ice)
Add 1ul of Plasmid
Incubate on Ice for 30min
Heat shock at 42Deg Cel. For 90sec
Incubate on Ice 2min
Add 450ul of LB medium and Incubate for 40-60min at 37 Deg Cel.
Seed 100ul on an Agar plate with the correct Antibiotics
Put at 37 Deg Cel over night.
Plates could be stored for 1 month sealed at 4 Deg Cel.