Samples were incubated in 9 volumes of buffered peptone water (BPW) (Oxoid, Basingstoke, U.K.) at 35°C for 18 hr. Aliquots of each BPW culture were then centrifuged at 13,800 × g for 5 min, and the supernatant was discarded. The pellet was resuspended in normal saline (1 ml) and centrifuged at 13,800 × g for 5 min. The supernatant was discarded, and the pellet was resuspended in 1 ml of 0.1× TE buffer with 5% Chelex 100 (Bio-Rad Laboratories, Hercules, CA, U.S.A.) before being boiled for 10 min, and then centrifuged at 13,800 × g for 10 min. The supernatant was used as a template for a PCR assay to detect E. albertii using primer pair lysP107F/lysP358R, as described previously [6]
Samples were incubated in 9 volumes of buffered peptone water (BPW) (Oxoid, Basingstoke, U.K.) at 35°C for 18 hr. Aliquots of each BPW culture were then centrifuged at 13,800 × g for 5 min, and the supernatant was discarded. The pellet was resuspended in normal saline (1 ml) and centrifuged at 13,800 × g for 5 min. The supernatant was discarded, and the pellet was resuspended in 1 ml of 0.1× TE buffer with 5% Chelex 100 (Bio-Rad Laboratories, Hercules, CA, U.S.A.) before being boiled for 10 min, and then centrifuged at 13,800 × g for 10 min. The supernatant was used as a template for a PCR assay to detect E. albertii using primer pair lysP107F/lysP358R, as described previously [6]
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