hygiena KIT230123 Spoilage Yeast Detection 1 LyoKit User Guide

hygiena KIT230123 Spoilage Yeast Detection 1 LyoKit

Specifications

• Product Name: Spoilage Yeast Detection 1 LyoKit
• Product No.: KIT230123 (DP)
• Revision: A, November 2023
• Application: PCR kit for qualitative or quantitative detection of Dekkera/Brettanomyces spp., Zygosaccharomyces spp., and Saccharomyces spp. using real-time PCR instruments

Product Usage Instructions

Program Setup:

1. Program your real-time PCR instrument with the following channels: FAM (Dekkera/Brettanomyces spp.), HEX (Zygosaccharomyces spp.), ROX (Saccharomyces spp.), and ATTO 490LS (Internal Control).
2. Set the program cycle as follows:

• 4 min | 5 min – 1 cycle
• 10 sec | 60 sec – 50 cycles (Fluorescence detection)

Data Interpretation:

Verify results of positive (Control Template) and negative (H2O) controls before interpreting sample results. Always compare samples to controls. Interpret results based on the table provided in the manual.

Preparation of the PCR Mix:

1. Place strips in rack and gently tap to move lyophilized pellets to the bottom of tubes.
2. Decap tubes immediately before filling with samples.
3. Add samples and controls to the tubes.
4. Seal tubes with provided 8-cap strips.
5. Mix thoroughly to resuspend pellet.
6. Centrifuge briefly, e.g., 5 seconds at 500 – 1,000 x g.
7. Start real-time PCR run by cycling samples as programmed.

FAQ

Q: What should I do if my sample results do not match the interpretation table?

A: In case of discrepancies, recheck your controls and ensure proper setup of the PCR instrument. If issues persist, contact customer support for assistance.

Q: Can this kit be used for other yeast species detection?

A: This kit is specifically designed for the detection of Dekkera/Brettanomyces spp., Zygosaccharomyces spp., and Saccharomyces spp. It is not validated for other yeast species detection.

Revision A, November 2023
Product No. KIT230123 (DP)
PCR kit for the qualitative or quantitative detection of Dekkera/Brettanomyces spp., Zygosaccharomyces spp. and Saccharomyces spp. 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 (Dekkera/Brettanomyces spp.), HEX (Zygosaccharomyces spp.), ROX (Saccharomyces spp.) and ATTO 490LS (Internal Control).

For the Dualo 32® Beverage 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 (H2O) controls, before interpreting the sample results. Always compare samples to positive and negative controls. Review data from each channel and interpret results as described in the table.

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 fi lter 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 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 pellet after sealing by mixing thoroughly.
   Alternatively, resuspend pellet by pipetting up and down multiple times in Step 3.

6. CENTRIFUGE
   Briefl y 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® Spoilage Yeast Detection 1 LyoKit

KIT230123
Kit for 48 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

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