hygiena KIT230131 LP Salmonella Plus Cronobacter Detection LyoKit User Guide

hygiena KIT230131 LP Salmonella Plus Cronobacter Detection LyoKit User Guide

Ready Reference Guide

Revision A, November 2023
Product No. KIT230131 (LP), KIT230132 (RP), KIT230133 (DP)

PCR kit for the qualitative detection of Salmonella and Cronobacter DNA using real-time PCR instruments.
Before starting, it is strongly recommended to read the entire product manual available on our website.

PROGRAM SETUP

Program your real-time PCR instrument before setting up the PCR reactions. Select the following channels:

• FAM (Salmonella), HEX (Cronobacter) and ROX (Internal Control).

For the BAX System Q7 instrument, no program setup is necessary.

Pre-incubation: 1 cycle
Step 1: 37 °C for 4 min
Step 2: 95 °C for 5 min
Amplifi cation: 50 cycles
Step 1 : 95 °C for 5 sec
*Step 2:** 60 °C for 60 sec

For some real-time PCR instruments the probe quencher as well as the usage of a passive reference dye has to be specified. This kit contains probes with a non-fluorescent “dark” quencher and no passive reference dye.
A Color Compensation is necessary for users of the LightCycler® 480 System: Color Compensation Set 5 (Product No. KIT230011).
For the Dualo 32® R2 real-time PCR instrument, please open the software, click on “New”, and select the respective template file. Template files can be added by clicking on “Add” in the “Select template file” window.
Product No. KIT230131 (LP), KIT230132 (RP), KIT230133 (DP)
Revision A, November 2023

DATA INTERPRETATION

Verify results of positive (Control Template) and negative controls (H2 O), before interpreting sample results.

Always compare samples to positive and negative control. Review results from each channel and for each sample. Ensure that each positive result has an amplification curve. In case no amplification curve is observed, the result is considered negative. Interpret results as described in the table.

Note: For interpretation of data generated with the BAX System Q7 instrument, please refer to the product instructions available on our website.

foodproof® is a registered trademark of Hygiena. The above mentioned real-time PCR instruments are registered trademarks of their respective holders.

PREPARATION OF THE PCR MIX

Take appropriate precautions to prevent contamination, e.g., by using filter tips and wearing gloves.

1. PLACE STRIPS IN RACK
   Take needed number of PCR tube strips out of aluminum bag.
   Important: close bag tightly afterwards. Place strips in a suitable PCR tube rack.
   If needed, gently tap the tubes to move the lyophilized pellets to the bottom of all tubes.
   

2. DECAP
   Immediately before filling, carefully open strips and discard caps.
   Do not leave open longer than necessary.
   

3. ADD SAMPLES AND CONTROLS
   Pipette 25 µL of DNA extract from foodproof lysis kits or 30 µL of DNA extract from BAXPrep lysis kit and 25 µL of Negative Control (colorless cap) and 25 µL of Control Template (purple cap) into respective wells.
   If using less volume, add PCR-grade H2 O to reach 25 µL or 30 µL, respectively.
   

4. SEAL
   Carefully seal the tubes with the provided 8-cap strips.
   

5. MIX
   Resuspend pellet after sealing by mixing thoroughly.
   Alternatively, resuspend pellet by pipetting up and down multiple times in Step 3.
   

6. CENTRIFUGE
   Briefly spin strips, e.g., 5 seconds at 500 – 1,000 x g, in a suitable centrifuge.
   

7. START REAL-TIME PCR RUN
   Cycle samples as described above.
   Place tubes in a vertical, balanced order into the cycler, e.g., two strips can be placed in the first and last column.
   

foodproof® Salmonella
plus Cronobacter
Detection LyoKit
KIT230131 /32 /33
Kit for 96 reactions
Store kit at 2 to 8 °C
For food testing purposes
FOR IN VITRO USE ONLY
Made in Germany
Hygiena® | Camarillo, CA 93012 USA | diagnostics.support@hygiena.com | www.hygiena.com

References

• Home | Hygiena

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