Fig 1: Evaluations on multiplex PCR. (a) Specificity evaluation of multiplex PCR. Lane M, DL2000 DNA molecular weight marker; lane S1, extracted genomic DNA mixture of reference strain (Table 4); lane S2, Actinobacillus pleuropneumoniae L20; lane S3, G. parasuis NO635; lane S4, Pasteurella multocida CGKV-VM02; lane S5, Streptococcus suis HB27; lane S6, Mycoplasma hyopneumoniae HN17; lane S7, enterotoxigenic Escherichia coli DDH135; lane S8, Salmonella Typhimurium CVCC542; lane S9, Staphylococcus aureus ATCC 25923; lane H2O, negative control. (b) Comparison of between previously reported single-plex PCR and our multiplex PCR. Sample types are as follows: 1, samples carrying S. suis; 2, carrying A. pleuropneumoniae and S. suis; 3, carrying G. parasuis and S. suis; 4, carrying A. pleuropneumoniae, G. parasuis, and S. suis; 5, carrying G. parasuis; 6, carrying A. pleuropneumoniae and G. parasuis.
Fig 2: Substrate specificity of the PhiKo endolysin. Chloroform permeabilized Gram-negative bacteria Thermus thermophilus HB8, T. thermophilus HB27, Thermus flavus ATCC 1087, Thermus parvatiensis DSM 21745, T. scotoductus MAT2631, and Gram-positive Deinococcus radiodurans ATCC 13939 were prepared and used as a substrate in turbidity reduction assay. The 100% activity corresponds to the highest decrease obtained among the dataset. Experiments were performed in triplicate; error bars indicate standard deviations.
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