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SUBMITTED 050113, REVISED 102513
Pick an isolated colony from non-selective or selective/differential agar medium with an inoculating loop
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or needle and resuspend (vortex or otherwise thoroughly mix) in 0.5 mL PBS in a sterile, capped tube.
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E.
Test Procedure
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General Preparation:
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(a)
This assay should be performed in a controlled laboratory environment.
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(b)
Do not use culture media or ANSR reagents beyond their expiration dates. Do not interchange
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reagents between ANSR kit lots.
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(c)
Remove ANSR reaction tubes from the foil pouch just before use. Avoid prolonged exposure to
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light. Tap reaction tubes on bench top to make sure that lyophilized reagents are at the bottom of the
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tube prior to adding the lysed sample.
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(d)
Complete all assay steps in sequence, avoiding delays between steps.
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(e)
Exercise care in pipetting steps to avoid cross-contamination of samples.
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(f)
Do not remove caps from reaction tubes at any point after the assay is started
; this will
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prevent accidental contamination of the environment with amplification products.
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(g)
Prior to starting the assay:
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(i)
Preheat the lysis heater block to 80 ± 2
o
C.
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(ii)
Start the ANSR software using the computer connected to the ANSR reader. Select “
Salmonella
”
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as the test type. Enter sample identifications and other experiment information. The reader will
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preheat to 56 ± 1
o
C.
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Assay Procedure:
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(a)
Add 50 µL of colony resuspension to a 1.2 mL cluster tube.
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(b)
Add 450 µL lysis buffer to the cluster tube. Note: It is not necessary to use the “lysis reagent”
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provided with the test kit for this application.
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