Chapter III. The BAX® System X5 PCR Assays
BAX® System X5 User Guide
45
Raw beef (including
seasoned and
frozen) in MP media
Homogenize 25 g sample in 225 mL pre-warmed
(42°C) BAX® System MP media. Incubate at 42°C for
9-24 hours.
None
Other raw meat
Homogenize 25 g sample in 225 mL pre-warmed
(42°C) BAX® System MP media. Incubate at 42°C for
24 hours.
None
2. Prepare BAX® System Lysis Reagent
Note: Do not over-mix the lysis reagent, as this can cause the mixture to foam.
To create a full bottle of BAX® System lysis reagent (prepares about 60 samples)
1. Add
150 μL of protease to one 12-mL bottle of lysis buffer.
2. Cap the bottle and invert it several times to mix.
To create smaller volumes of BAX® System lysis reagent (prepares about 4 samples)
1. Add
12.5μL protease to 1 mL of lysis buffer in a separate sterile container.
2. Seal the container and invert it several times to mix.
3. Perform Sample Lysis
1. Make sure that the cooling blocks have been refrigerated or otherwise chilled to 2-8°C. If you find that they
have NOT been adequately chilled, call for technical assistance.
2. Create a rack file according to
the instructions found under “Creating a Rack File”.
3. Turn on the Automated Thermal Block and select the Gram Negative program.
Note: Lysis may also be performed using separate analog heating and cooling blocks. See
“Using Analog Heating and
Cooling Blocks with the BAX® System Assays
” for details and protocols.
4. Break cluster tubes apart. Label and arrange cluster tubes in the rack according to your rack file.
5. Transfer 200
μL lysis reagent to each of the cluster tubes.
6. Transfer 5
μL enriched sample to the corresponding cluster tubes, using a new pipette tip for each sample.
Note: Do not shake or mix the enrichment before transferring samples to cluster tubes. If samples have been agitated, let
sit for at least 10 minutes before transferring aliquots to cluster tubes.
7. After all transfers have been completed, secure the caps.
8.
After the “Load Samples” prompt appears on the Automated Thermal Block, place the rack of cluster tubes
on the Automated Thermal Block.
9. Press the SELECT/CONTINUE button to begin automated lysis.
Note: This process takes about 1 hour to complete.
10. When sample lysis is complete, the Automated Thermal Block displays the message
“Sample PCR Ready”
and sounds four audible beeps.
