hygiena KIT230081 LP Cronobacter Detection LyoKit User Guide

hygiena KIT230081 LP Cronobacter Detection LyoKit

Product Information

Specifications

• Product Name: Cronobacter Detection LyoKit
• Revision: A, November 2023
• Product Numbers: KIT230081 (LP), KIT230082 (RP), KIT230083 (DP)
• Type: PCR kit for qualitative detection of Cronobacter spp. DNA
• Usage: Real-time PCR instruments

Product Usage Instructions

Program Setup

Before starting, program your real-time PCR instrument with the following settings:

• Channels: FAM (Cronobacter spp.) and VIC (Internal Control)
• Cycling Conditions: 4 min at 5 sec | 60 sec for 1 cycle, followed by 50 cycles with fluorescence detection

Data Interpretation

To interpret results, verify positive and negative controls before analyzing sample results. Compare samples to controls and interpret based on the FAM and VIC channels according to the provided table.

Preparation of the PCR Mix

1. Place Strips in Rack: Remove PCR tube strips from the aluminum bag, close the bag tightly, and place the strips in a rack.
2. Decap: Open strips, discard caps, and avoid leaving them open for long.
3. Add Samples and Controls.
4. Seal: Carefully seal tubes with the provided caps.
5. Mix: Resuspend the pellet by mixing thoroughly or pipetting up and down.
6. Centrifuge: Spin strips briefly in a suitable centrifuge.
7. Start Real-time PCR Run: Cycle samples as instructed and place tubes in a balanced order in the cycler.

Note: Take precautions to prevent contamination by using filter tips and gloves during the process.

FAQ

Q: What should I do if my results are invalid?

• A: If your results are invalid, ensure proper handling of samples and controls, repeat the process with caution, and refer to the manual for troubleshooting tips.

Q: Can I reuse the PCR tube strips?

• A: It is recommended to use new PCR tube strips for each new set of samples to avoid cross-contamination.
• Product No.: KIT230081 (LP), KIT230082 (RP), KIT230083 (DP)
• PCR kit for the qualitative detection of Cronobacter spp. 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 (Cronobacter spp.) 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 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.

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® Cronobacter Detection LyoKit
• KIT230081 /82 /83 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

Documents / Resources

| hygiena KIT230081 LP Cronobacter Detection LyoKit [pdf] User Guide
KIT230081 LP, KIT230082 RP, KIT230083 DP, KIT230081 LP Cronobacter Detection LyoKit, Cronobacter Detection LyoKit, Detection LyoKit, LyoKit
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References

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