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5

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before working with potentially infectious materials. All enrichment broths

should be sterilized following any culture based confirmatory steps.

General Preparation

(a) 975 ± 19.5 mL of pre-warmed (41.5 ± 1° C) 3M ISO BPW enrichment

media

(b) Use aseptic techniques

(c) Use filter laboratory bags during enrichment to minimize particulates

(d) Separate work areas for the following: media preparation, sample

preparation, and pathogen detection

(e) Clean the work stations with a disinfectant of choice before and after use.

(Sodium hypochlorite solution, phenol solution, Quaternary ammonium

solution, etc.)

(f) Do not reuse kit disposables

(g) Change pipette tips in between samples

(h) Wear personal protective equipment (PPE)

DNA Lysis

(a) Allow the lysis solution tubes to warm to room temperature (20-25° C)

(b) Pre-warm dry bath incubator to 100 ± 1°C.

(c) Transfer enriched sample into an individual lysis tube.

DNA Amplification

(a) Use of aseptic technique

(b) Change pipette tips between samples

(c) Use gloves and protective laboratory wear

(d) Do not touch any molecular equipment and supplies without wearing

gloves

(e) Ensure that the reagent tubes are completely sealed with caps