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| - | {{Team:Alberta/beginMainContent}}
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| - | Protocol 1: Transformations
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| - | What you will need:
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| - | * Competent DH5α cells (50 uL per transformation)
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| - | * 2 uL of plasmid you want to transform
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| - | * 800 uL of LB (non-contaminated) per transformation
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| - | * Plate with correct antibiotics
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| - | Procedure:
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| - | * Chill a box of pipette tips at -4 C and put your reaction tubes on ice.
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| - | * Obtain DH5α cells from the –80 C freezer and thaw in palm. Just as they thaw, store on ice for 10 minutes.
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| - | * Use a chilled tip to transfer 50 ul of the DH5α cells into each tube. Set aside one tube as a control.
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| - | * Pipet 2uL of the plasmid (up to 25ng per 50 ul of competent cells, plasmid amt. should not exceed 5% that of the competent cells) into each tube. Swirl.
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| - | * Store on ice for 30 minutes.
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| - | * Place tubes in incubator set to 42 C for EXACTLY 90s.
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| - | * Return the tubes to ice for 2 min.
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| - | * Add 800uL of LB to each tube.
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| - | * Incubate at 37 C for 45 min, ensure that the tube has at least 2ml capacity, to properly aerate the cells.
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| - | * Spread cells on plate with appropriate antibiotic: plate between 50- 300ul each. Let dry.
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| - | * Place plates inverted in incubator at 37 C for 12-16 hours.
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| - | {{Team:Alberta/endMainContent}}
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| - | [[Team:Alberta/Protocols]]
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