DNA Effects on Transformation Efficiency and Colony Output: The optimal amount of DNA to use in a transformation reaction is lower than commonly recognized. Using clean, supercoiled pUC19, the efficiency of transformation is highest in the 100 pg-1 ng range. However, the total colonies which can be obtained from a single transformation reaction increase up to about 100 ng.Effect of heat shock time on NEB 5-alpha competent E.coli transformation efficiency: 50 μl of competent cells were transformed with 100 pg of pUC19 control DNA following the provided High Efficiency Transformation Protocol except heat shock time varied from 0 to 80 seconds.Effect of DNA incubation time on NEB 5-alpha competent E.coli transformation efficiency: 50 μl of competent cells were transformed with 100 pg of pUC19 control DNA following the provided High Efficiency Transformation Protocol except DNA incubation time varied from 0 to 40 minutes.Effect of outgrowth medium on transformation efficiency: 50 μl of NEB 5-alpha competent E.coli was transformed with 100 pg of pUC19 control DNA following the provided High Efficiency Transformation Protocol with the exception of varying the outgrowth medium. NEB SOC outgrowth medium delivers the highest transformation efficiency.Benefit from the high transformation efficiencies of NEB 5-alpha: The transformation efficiencies of NEB 5-alpha and DH5α were compared using each manufacturers' recommended protocols. Values shown are the average of triplicate experiments.
For C2987H, C2987I, C2987P and C2987U, SOC Outgrowth Medium is provided. For C2987R, Outgrowth Medium 1.5 is provided.
CAUTION: This product contains DMSO, a hazardous material. Review the MSDS before handling.
STORAGE AND HANDLING: Competent cells should be stored at -80°C. Storage at -20°C will result in a significant decrease in transformation efficiency. Cells lose efficiency whenever they are warmed above -80°C, even if they do not thaw.
SOC medium can be stored at either 4°C or Room Temperature depending on how fast it will be used. Storing at Room Temperature is convenient and adequate for short term usage (weeks to a couple of months). For long term storage, we recommend storing at 4°C. Please note that Outgrowth Medium 1.5 supplied with C2987R (1 x 384 well plate format) must only be stored at Room Temperature or crystals will form.
Kostylev M., Otwell A.E., Richardson R., Suzuki Y. (2015) Cloning Should Be Simple: Escherichia coli DH5α-Mediated Assembly of Multiple DNA Fragments with Short End Homologies PLoS One; 10, e0137466. PubMedID: PMC4562628, DOI: 10.1371/journal.pone.0137466
Lee S.B., Park S.K., Kim Y.S. (2015) Production of bioactive chicken (Gallus gallus) follistatin-type proteins in E. coli Sci Rep; 5, 11603. PubMedID: 26302688, DOI: 10.1038/srep11603
Mascher G, Derman Y, Kirk DG, Palonen E, Lindström M, Korkeala H (2014) The CLO3403/CLO3404 Two-Component System of Clostridium botulinum E1 Beluga Is Important for Cold Shock Response and Growth at Low Temperatures Appl Environ Microbiol; 80(1), 399-407. PubMedID: 24185852, DOI: 10.1128/AEM.03204-13
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