hygiena KIT230099 (LP) foodproof Salmonella Detection LyoKit User Guide

hygiena KIT230099 (LP) foolproof Salmonella Detection LyoKit

Product Information

Specifications

• Product Name: Salmonella Detection LyoKit
• Revision: A, November 2023
• Product Numbers: KIT230099 (LP), KIT230101 (RP), KIT230102 (DP), KIT230100 (LP – Large)
• PCR Kit for the qualitative detection of Salmonella DNA using real-time PCR instruments

Product Usage Instructions

Program Setup:

1. Program your real-time PCR instrument before setting up the PCR reactions.
2. Select the following channels: FAM (Salmonella) and VIC (Internal Control).
3. Set the program cycle as follows:
   • 4 min | 5 min – 1 cycle
   • 5 sec | 60 sec – 50 cycles (Fluorescence detection)

Preparation of the PCR Mix

1. Take appropriate precautions to prevent contamination by using filter tips and wearing gloves.
2. Place strips in a suitable PCR tube rack and tap gently to move lyophilized pellets to the bottom of all tubes.
3. Decap tubes immediately before filling, ensuring not to leave them open longer than necessary.
4. Add samples and controls to the tubes.
5. Seal the tubes with the provided 8-cap strips.
6. Mix thoroughly to resuspend pellets.
7. Briefly spin strips in a suitable centrifuge before starting the real-time PCR run.

FAQ

Q: Can I reuse the PCR tube strips?

• A: No, it is recommended to use new PCR tube strips for each PCR run to prevent potential contamination.

Q: How should I store the kit when not in use?

• A: Store the kit as per the storage instructions provided in the product manual to maintain its integrity and effectiveness.

Q: What should I do if I encounter an ‘Invalid’ result?

• A: If you receive an ‘Invalid’ result, review your process to ensure proper handling and follow the troubleshooting steps outlined in the manual.
• Product No. KIT230099 (LP), KIT230101 (RP), KIT230102 (DP), KIT230100 (LP – Large)
• PCR kit for the qualitative detection of Salmonella 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) and VIC (Internal Control).
• As an alternative to VIC, HEX can be used. For the PikoReal® 24, Yakima Yellow has to be selected.
• Pre-incubation: 1 cycle
  • Step 1: 37 °C for 4 min
  • Step 2: 95 °C for 5 min
• Amplification: 50 cycles
  • Step 1: 95 °C for 5 sec
  • Step 2: 60 °C for 60 sec
• Fluorescence detection
• 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.
• For the Dualo 32 R2 and the Dualo 32 Beverage real-time PCR instruments, 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.

DATA INTERPRETATION

• Verify results of positive (Control Template) and negative controls (H2O), before interpreting sample results.
• Always compare samples to positive and negative control.
• Review data from each channel and interpret results as described in the table.

PREPARATION OF THE PCR MIX

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

1. PLACE STRIPS IN THE RACK

   • Take the needed number of PCR tube strips out of an aluminum bag.
   • Important: close the bag tightly afterward. 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 it open longer than necessary.

3. ADD SAMPLES AND CONTROLS

   • Pipette 25 µL of samples, Negative Control (colorless cap), or Control Template (purple cap) into respective wells.
   • If using less volume, add PCR-grade H2O to reach 25 µL.

4. SEAL

   • Carefully seal the tubes with the provided 8-cap strips.

5. MIX

   • Resuspend the pellet after sealing by mixing thoroughly.
     Alternatively, resuspend the 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 Detection LyoKit
• KIT230099 /100 /101 /102 Kit for 96 (480) 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

Documents / Resources

| hygiena KIT230099 (LP) foodproof Salmonella Detection LyoKit [pdf] User Guide
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References

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