hygiena KIT230048 Listeria monocytogenes Detection Kit User Guide
- June 20, 2024
- Hygiena
Table of Contents
foodproof®
Listeria monocytogenes Detection Kit
Ready Reference Guide
Revision A, December 2023
Product No. KIT230048
PCR kit for the qualitative detection of L. monocytogenes 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 (L. monocytogenes) 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
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 usage of a passive reference dye has to be specified. This kit contains probes with TAMRA as quencher and no passive reference dye.
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.
| FAM | VIC | Result Interpretation |
|---|---|---|
| + | + or – | Positive for L. monocytogenes |
| – | + | Negative for L. monocytogenes |
| – | – | Invalid |
PREPARATION OF THE PCR MIX
Take appropriate precautions to prevent contamination, e.g., by using fi lter
tips and wearing gloves.
Thaw reagents, mix (do not vortex!) and briefl y spin vials before opening.
-
PREPARE PCR MIX
Add 18 µL of Master Mix (yellow cap), 1 µL of Enzyme Solution (red cap) and 1 µL of Internal Control (white cap) for each reaction to a suitable tube (n samples + 2 controls + at least one additional reaction to cover pipetting loss).
Mix carefully but thoroughly by pipetting up and down. -
ADD PCR MIX
Pipet 20 µL of prepared PCR mix into each strip or plate well. -
ADD SAMPLES AND CONTROLS
Pipet 5 µL of samples, Negative Control (colorless cap) or Control Template (purple cap) into respective wells. -
SEAL
Seal strips/plate accurately. -
CENTRIFUGE
Briefl y spin strips/plate in a suitable centrifuge. -
START REAL-TIME PCR RUN
Cycle samples as described above.
foodproof®
Listeria monocytogenesDetection Kit
KIT230048
Kit for 96 reactions
Store kit at -15 to -25 °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) >>