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XL10-Gold Cells Supplied in New Single-Reaction Format

e SoloPack single-tube, single-reaction format is available at the supercompetent efficiency of 1 x 109 transformants/g of supercoiled DNA and the competent efficiency of 1 x 108 transformants/g of supercoiled DNA.

Although this new format is convenient, it is especially important to maintain high transformation efficiency and not compromise the outcome of the cloning experiment. Hence, we chose our most efficient competent cell strain, XL10-Gold, and created SoloPack Gold supercompetent cells and SoloPack Gold competent cells. Use the supercompetent efficiency (1 x 109 transformants/g of supercoiled DNA) for PCR cloning, restriction enzyme cloning, the ligase-independent cloning (LIC) method,3 and expression vector construction. Or, as an economical alternative, use the competent efficiency of 1 x 108 transformants/g of supercoiled DNA when high efficiency is not required. For plasmid library construction and other demanding experiments where optimal transformation efficiency is critical, Stratagene provides the XL10-Gold ultracompetent cells in our standard-tube format with efficiency greater than 5 x 109 transformants/g of supercoiled DNA.

Transformation

We tested the transformation efficiency of 1 x 109 transformants/g of supercoiled DNA for the SoloPack Gold supercompetent cells: The cells were thawed on ice, 1 l of XL10-Gold b-mercaptoethanol mix was added, and the cells were incubated on ice for 10 minutes. Next, 10 pg of pUC DNA was added to the cells and they were incubated on ice for an additional 20 minutes. The cells were then heat pulsed at different temperatures for varying lengths of time to optimize efficiency. After the heat pulse, 0.15 ml of NZY+ outgrowth media was added, and the cells were shaken in the reaction tubes for 60 minutes prior to plating. Our results showed t
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