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E. coli (DE3) containing pET28-LbCas12a-WT or pET28-LbCas12a-D1180A were grown in LB broth until an OD600 of 0.6 was achieved. Recombinant LbCas12a protein expression was induced by adding 0.8 mM IPTG and incubating for 16 h at 16 °C. The E. coli was lysed using a buffer conta...
The cells were cultured at 37 °C with shaking at 300 rpm in 1 L of LB broth containing kanamycin (30μg mL− 1) until an OD600 of 0.6–0.8 was reached. Expression of the recombinant protein was induced by adding 0.4 mM IPTG, followed by cultivation for 16 h at 25 °C and 200...
monocytogenes were grown in Brain Heart Infusion (BHI) broth to an OD600 of 0.80, and 1 ml of each culture was taken for each binding reaction. Bacterial cells were washed twice in buffer with 20 mM HEPES pH 7.5, 150 mM NaCl, resuspended in 1 ml of binding buffer (20 mM HEPES pH ...
2.3. Quantitative estimation of GABA in fermented broth Each tested LAB strain was transferred to MRS liquid medium after activation and incubated at 37 °C for 24 h to logarithmic period. Then, the strains were inoculated at 4% inoculum amount into a fermentation medium and incubated at 37 °...