CaCl2 Competent Cells


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Transformation protocol

Melt agar in autoclave or microwave. Cool down to 60 oC. One jar ( 100mL ) is enough for 7 plates.

-Add 10 μl of ligation product to 100 μl of cells ( don't for get controls! )

-Incubate 30 min on ice (make agar plates)

-Incubate for 1 min at 42 oC

-Add 400 μl of TY

-Incubate for 30 min at 37 oC (dry agar plates)

-Plate out. Use 250 μl of the transformed cells.

  • If you expect the transformation to be very efficient (for instance using undigested plasmid DNA) first make dilution (1:10 or 1:100 ) and plate 250 μl of this
  • If you expect the trasformation to be inefficient, concentrate cells by spinning the 500 μl and resuspending the pellet in 250 μl, which you will use for plating.