hygiena KIT230121 LP Spoilage Yeast Detection 1 LyoKit User Guide
- May 15, 2024
- Hygiena
Table of Contents
foodproof®
Spoilage Yeast Detection 1 LyoKit
Ready Reference Guide
Revision A, November 2023
Product No. KIT230121 (LP), KIT230122 (RP)
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 Cy5 (Internal Control).
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
- Fluorescence detection
For some real-time PCR instruments the probe quencher as well as the use of a
passive reference dye must 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).
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.
| FAM + | HEX | ROX | Cy5 | Result Interpretation |
|---|---|---|---|---|
| + | + or – | + or – | + or – | Positive for Dekkera/Brettanomyces spp. |
| + or – | + | + or – | + or – | Positive for Zygosaccharomyces spp. |
| + or – | + or – | + | + or – | Positive for Saccharomyces spp. |
| – | – | – | + | Negative for targeted spoilage yeasts |
| – | – | – | – | Invalid |
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.
-
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
Briefl y 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 1 LyoKit| KIT230121 / 22
Kit for 48 reactions
Store kit at 2 to 8 °C| For food testing purposes
FOR IN VITRO USE ONLY
Made in Germany
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Hygiena®
Camarillo, CA 93012 USA
[email protected]
www.hygiena.com
Documents / Resources
|
hygiena KIT230121 LP Spoilage Yeast Detection 1
LyoKit
[pdf] User Guide
KIT230121 LP, KIT230121 LP Spoilage Yeast Detection 1 LyoKit, Spoilage Yeast
Detection 1 LyoKit, Yeast Detection 1 LyoKit, Detection 1 LyoKit, 1 LyoKit
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