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CaCl2 Competent Cells
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(New page: 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...)
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(New page: 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...)
Newer edit →
Revision as of 10:28, 24 August 2010
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.
