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Strain, Selection plate
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QuickStep-Cloning
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RF Cloning
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Transformation efficiency [cfu/μg]
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|---|
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DH5α, Ampicillin
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0
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0
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3.8 · 104
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DH5α, Kanamycin
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476
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35
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3.8 · 104
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C41(DE3), Kan + IPTG
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618(575)
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160(7)
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4.2 · 106
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- Colony counts for E. coli strains DH5α and C41 (DE3) transformed with the products of RF cloning and of QuickStep-Cloning and plated on agar plates supplemented with: (i) 100 μg/ml ampicillin, (ii) 50 μg/ml kanamycin, and (iii) 50 μg/ml kanamycin and 1 mM IPTG. Transformation efficiency was determined based on concurrent transformation of 1 ng intact pET24a-HLTEV-p53 plasmid. Numbers in the brackets denote EGFP-expressing colonies, as determined by visual inspection using UV transilluminator. Lack of colonies observed on ampicillin-supplemented agar plates indicated that the final PCR mixture produced via QuickStep-Cloning, used directly for bacterial transformation, did not contain significant amount of donor plasmid
