hygiena KIT230124 Spoilage Yeast Detection 2 LyoKit User Guide
- May 15, 2024
- Hygiena
Table of Contents
hygiene KIT230124 Spoilage Yeast Detection 2 LyoKit
Product No. KIT230124 (LP), KIT230125 (RP)
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 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
Melting Curve: 1 cycle
- Step 1: 95 °C for 50 sec
- Step 2: 50 °C for 50 sec
- Step 3**: ramp up to 85 °C
- Fluorescence detection
- Fluorescence detection during 50 – 85 °C
- ramp with 1 – 2 measurements/°C
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). Product No. KIT230124 (LP), KIT230125 (RP) Revision A, November 2023
DATA INTERPRETATION
Verify the results of positive (Control Template) and negative (H2 O) 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.
PREPARATION OF THE PCR MIX
Take appropriate precautions to prevent contamination, e.g., by using
filter tips and wearing gloves.
PLACE STRIPS IN THE RACK
Take the needed number of PCR tube strips out of the aluminium bag.
Important : close the 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 filling, carefully open strips and discard caps. Do not
leave it open longer than necessary.
ADD SAMPLES AND CONTROLS
Pipette 25 µL of samples, Negative Control (colourless cap) or Control Template (purple cap) into respective wells. If using less volume, add PCR- grade H2 O to reach 25 µL.
SEAL
Carefully seal the tubes with the provided 8-cap strips.
MIX
Resuspend the pellet after sealing by mixing thoroughly. Alternatively,
resuspend the 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 first and last column.
foodproof® Spoilage
Yeast Detection 2 LyoKit
KIT230124 / 25
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) >>