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Highest Possible Transformation Efficiencies for High-Throughput ,,, Applications

of plasmid miniprep DNA. These cells are recombination-deficient (recA), ensuring the stability of inserted DNA. With the lacIqZDM15 gene on the F episome, 96Pack Gold competent cells allow bluewhite color screening of recombinant plasmids. In addition, this cell strain is restriction-minus, a feature that increases the efficiency of introducing eukaryotic DNA into E. coli and increases the size and representation of libraries constructed with methylated or hemimethylated DNA. Finally, 96Pack Gold cells grow at a fast rate, resulting in the formation of larger, easier-to-select colonies (Figure 1).

Superior Performance

We compared the transformation efficiency of 96Pack Gold cells with MultiShot cells (Invitrogen) (Figure 2). To test efficiency with supercoiled DNA, we added 1 pg and 10 pg of the pUC plasmid to either 96Pack Gold or MultiShot cells. To test efficiency for ligated DNA, we transformed 96Pack Gold cells and MultiShot cells with 2.3 ng of a ligated construct, the pCMV-Script plasmid with the LacZ gene insert (approximately 4.9 kb). For each experiment, we followed the manufacturers protocols. As can be seen, 96Pack Gold competent cell transformation efficiency outperformed the competitors product with both supercoiled and ligated DNA.

Conclusions

Stratagenes high-efficiency 96Pack Gold competent cells, provided in a high-throughput format, are the superior choice for PCR cloning, restriction enzyme cloning, and ligase-independent cloning. In comparison transformations of either s
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