The total daily amount of bacteria in the probiotic capsule was 8-9 × 109 colony forming units, with an equal amount of each strain.
Each microarray hybridization reaction consisted of genomic DNAs from the reference strains (an equal amount of DNA from C. jejuni strain NCTC11168 and strain RM1221) and from a test strain that were fluorescently labeled with indodicarbocyanine (Cy5) and indocarbocyanine (Cy3), respectively.
Each of the three knockdown mutants and the two reference strains were grown separately before equal amounts of each strain were mixed and used to inoculate (i) 7H9 medium, (ii) Sauton's medium with glycerol as the primary carbon source, and (iii) Sauton's medium with butyrate as the primary carbon source.
In cases of mice inoculated with two strains, the 0.5 ml suspension contained equal amounts of each strain (final concentration, 2×109 cfu per ml).
10 libraries were mixed in a proportion to maintain an equal amount of DNA from each strain in the multiplexed sequencing library.
For phenotype assessment, ΔCYC2 strains were starved for 24 h and mixed with equal amount of each mating type of strains to induce meiosis.
Total proteins of each strain were extracted from the lysed cells and equal amount of protein extracts from each strain were labeled with Cy2, Cy3, or Cy5 (GE Healthcare, Piscataway, NJ), respectively.
Equal amount of collected mycelia from each strain was transferred to the same media freshly prepared, for another 0, 6, 12, 24, 36, 48, 60, 72, and 84 h, respectively.
In order to detect gene expressions by both R265 and R272 strains, a reference cDNA pool was prepared by mixing together an equal amount of RNA obtained from both strains after growth for 1 hr in eight different growth conditions based on previous publication [36].
As a control, an equal amount of extract from a non-hemolytic strain of GBS (Δ cylE) was included.
