( higher than 5 x 10 9 tra...)Highest Possible Transformation Efficiencies for High-Throughput ,,, Applications

higher than 5 x 10⁹ transformants per microgram of supercoiled pUC18 vector (2.7 kb); efficiencies for ligated DNA were lower as expected. With the advent of the Hte phenotype, transformation efficiencies for ligated DNA increased 25-fold. This phenotype allows competent cells to better accept ligated or larger DNA plasmids, which are known to transform less efficiently than small plasmids on a per molecule basis. Due to exceptional performance, we have found that competent cells with the Hte phenotype are the best choice for any cloning experiment that demands a high efficiency of retrieving transformants from ligation reactions, such as constructing plasmid libraries and PCR cloning. The Hte phenotype helps 96Pack Gold cells fulfill the need for streamlined transformation procedures and superior performance.

In the basic high-throughput procedure, DNA is added to each well of 96Pack Gold cells with a multichannel pipettor. The microtiter plate is chilled and then subjected to heat shock, which is simply performed with either a thermocycler, a heating block, or a conventional water bath. Growth medium is added to the wells. After incubations, transformants are selected with the appropriate antibiotic medium. This simple protocol saves time and reagents, while providing the highest possible transformation efficiencies.

Engineered to Achieve the Highest Performance

96Pack Gold competent cells carry the Hte mutation, which confers the capacity to transform large and ligated DNA molecules at a higher efficiency. The 96Pack Gold strain is endonuclease-deficient (endA1), which greatly improves the quality
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