After adding 6 mL's of Detection Buffer mixture solution to each sample, squeeze the sponge at least 3 times and ensure the buffer mixture is readsorbed into the sponge. Roll the bags and incubate at the specified temperature 35 +/- 1°C for 6 hours +/- 1 hour.
**For most accurate results incubate as close to 6 hours as possible**
The incubation period for the DETECT/L method is not considered an enrichment. The 6 mL's of buffer solution mix is intended to counter any sanitizer in the sponge. The incubation also allows any Listeria cells to recover so that the bacteriophage can seek out the Listeria cells and utilize the cells to produce the reporter enzyme, half of the reaction responsible for light production (and Listeria detection!)