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The resultant stock solution was diluted appropriately to 1 ml to achieve a countable dilution of cells per square of the cell counter.
The application of nested PCR technique in this study was able to identify V. harveyi strains at serial dilution of cells density as low as 100 cfu/mL, which is equal to a single cell or at DNA concentration up to 101 fg/µL.
At least 6 wells were examined for every dilution of cells.
Titration curves made with serial dilution of cells were not modified by addition of 1-mT, demonstrating that the compound does not influence the assay.
In total, six clones were selected by the serial dilution of cells.
The next day, cell concentrations were determined, and an initial dilution of cells to 2.4 × 10 cells/ml was made.
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(A) A serial dilution of cell culture spotted onto solid medium containing 100 µg/mL ampicillin and 5 ng/mL ciprofloxacin.
The titers of intracellular bacteria were determined by serial dilution of cell lysates on agar plates.
A 1 10 (v/v) dilution of cell suspension and ViaCount reagent (Guava Technologies) was prepared for each sample.
Then, 250 μl of each dilution of cell culture was mixed with 250 μl of the broth-faecal suspension and 500 μl of BHI in a new eppendorf tube.
Beads were washed twice with washing buffer (1/3 dilution of cell lysis buffer in phosphate-buffered saline (PBS)), and briefly centrifuged and resuspended in the sample buffer before being subjected to SDS-PAGE.
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