hygiena KIT230126 Spoilage Yeast Detection 2 LyoKit User Guide
- May 15, 2024
- Hygiena
Table of Contents
foodproof®
Spoilage Yeast Detection 2 LyoKit
Ready Reference Guide
Revision A, November 2023
KIT230126 Spoilage Yeast Detection 2 LyoKit
Product No. KIT230126 (DP)
PCR kit for the qualitative detection of Saccharomyces cerevisiae var.
diastaticus, Wickerhamomyces anomalus, Kazachstania exigua and
Schizosaccharomyces pombe 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 (Saccharomyces cerevisiae var. diastaticus), HEX (Wickerhamomyces
anomalus),
ROX (Kazachstania exigua and Schizosaccharomyces pombe) and ATTO 490LS
(Internal Control).
Fluorescence detection
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 10 sec
Step 2: 60 °C for 60 sec
Melting Curve: 1 cycle
Step 1 : 95 °C for 50 sec
Step 2 : 37 °C for 50 sec
Step 3: ramp up to 85 °C
For the Dualo 32® (for beverage testing) 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 (H2 O),
before interpreting sample results.
Always compare samples to positive and negative controls. Review data from
each channel and interpret results as described in the table.
| FAM | HEX | ROX | ATTO 490LS | Result Interpretation |
|---|---|---|---|---|
| + | + or – | + or – | + or – | Positive for Saccharomyces cerevisiae var. |
diastaticus
+ or –| +| + or –| + or –| Positive for Wickerhamomyces anomalus
+ or –| + or –| +| + or –| Positive for K. exigua and Schizosaccharomyces
pombe
–| –| –| +| Negative for targeted spoilage yeasts
–| –| –| –| Invalid
Melting Curve| Channel| Organism| Tm-Range
---|---|---|---
ROX| Kazachstania exigua| 68 ± 2°C
Schizosaccharomyces pombe| 78 ± 2°C
PREPARATION OF THE PCR MIX
Take appropriate precautions to prevent contamination, e.g., by using fi lter tips and wearing gloves.
-
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. -
DECAP
Immediately before fi lling, carefully open strips and discard caps.
Do not leave open longer than necessary. -
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. -
SEAL
Carefully seal the tubes with the provided 8-cap strips. -
MIX
Resuspend pellet after sealing by mixing thoroughly.
Alternatively, resuspend pellet by pipetting up and down multiple times in Step 3. -
CENTRIFUGE
Briefly spin strips, e.g., 5 seconds at 500 – 1,000 x g, in a suitable centrifuge. -
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 fi rst and last column.
foodproof® Spoilage
Yeast Detection 2 LyoKit
KIT230126
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
Read User Manual Online (PDF format)
Read User Manual Online (PDF format) >>