PROTOCOLS
6.TRANSFORMATION
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1.Sterilized the enviroment.
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2.Take the compatent cells from -80 ºC thawing on ice.
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3.Add 50 µL of thawed competent cells into 1.5ml tube.
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4.Add 1 - 2 µL of the resuspended DNA to the 2ml tube. Pipet up and down a few times, gently.
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5.Centrifuge tubes at 3000 rpm for 30-45 seconds.
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6.Incubate the cells on ice for 45 minutes.
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7.Heat the tubes in the 42 C heat block for a 80 seconds.
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8.Incubate the cells on ice for 5 minutes.
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9.Add 450 μl of LB broth (make sure that the broth does not contain antibiotics and is not contaminated) to each transformation
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10.Incubate the cells at 37ºC for 1 hour while the tubes are shaking.
11.Label petri dish with LB agar and the appropriate antibiotic.
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12.Plate 135 µl of the transformation onto the dishes, and spread.
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13.Incubate the plates at 37ºC for 12-14 hours, making sure the agar side of the plate is up.
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