pET26-b-PlpE

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	The ligation reaction was transformed into competent E. coli BL21 (DE3), and the transformed cells were selected on TSA plates containing 50 µg/ml kanamycin.

Protocol tips
The ligation reaction was transformed into competent E. coli BL21 (DE3), and the transformed cells were selected on TSA plates containing 50 µg/ml kanamycin.

Publication protocol

The ligation reaction was transformed into competent E. coli BL21 (DE3), and the transformed cells were selected on TSA plates containing 50 µg/ml kanamycin. The selected clones were further analyzed by PCR using specific and also universal T7 primers. Finally, the recombinant plasmid of two positive clones was sequenced using T7 promoter and terminator primers. Five ml of TSA medium with 50 µg/ml kanamycin was inoculated with a fresh colony of recombinant clone and incubated at 37 °C with shaking. When the optical density of the culture at a wavelength of 600 nm reached 0.6, protein expression was induced with a final concentration of 1mM isopropyl-D-1- thiogalactopyranoside (IPTG).

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