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| - | {{Caltech_iGEM_10|
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| - | Content=
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| - | __NOTOC__
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| - | ==Making Competent Cells==
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| - | ===Materials===
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| - | * N samples of DNA to be transformed, 1 positive control sample
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| - | * N+1 electroporation cuvettes
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| - | * N+1 50μL aliquots of EC cells (we used DH5α)
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| - | * N+1 mL SOC (Super-optimal broth + 10mM glucose)
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| - | * N+1 LB-agar plates of appropriate resistances
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| - | ===Procedure===
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| - | # Chill electroporation cuvettes and thawed EC cells on ice. Do not let EC cells warm to above ice-cold.
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| - | # Add 1μL of ligation product to the 50μL aliquot. Don't forget an aliquot for positive control.
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| - | # Transfer DNA/cell mixture to a cuvette and pulse at 2.5V. Rescue the cells immediately by adding 0.75mL warm SOC to the cuvette.
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| - | ## Ensure that the time constant is above 3.0 for each.
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| - | # Incubate for 1 hour at 37°C.
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| - | # Plate the entire mixture on an LB-agar plate with the appropriate antibiotic resistance, grow overnight.
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| - | }}
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Latest revision as of 23:42, 14 October 2010
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