hygiena KIT230106 LP Magnetic Preparation Kit User Guide
- June 17, 2024
- Hygiena
Table of Contents
foodproof®
Salmonella Enteritidis & Typhimurium
Detection LyoKit
Ready Reference Guide
Product No. KIT230106 (LP), KIT230107 (RP)
Revision A, November 2023
PCR kit for the qualitative detection of Salmonella Enteritidis and Salmonella
Typhimurium 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 (Salmonella Enteritidis), HEX (Salmonella Typhimurium) and ROX (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 3 (Product No. KIT230005).
For colony confirmation, a shortened PCR protocol is available. Please refer
to the manual.
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 | Result Interpretation |
|---|---|---|---|
| + | + | + or – | Positive for Salmonella Enteritidis and Salmonella Typhimurium |
| – | + | + or – | Positive for Salmonella Typhimurium |
| + | – | + or – | Positive for Salmonella Enteritidis |
| – | – | + | Negative for Salmonella Enteritidis and Salmonella Typhimurium |
| – | – | – | Invalid |
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® Salmonella
Enteritidis & TyphimuriumDetection LyoKit
KIT230106 / 07
Kit for 96 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 | [email protected] | www.hygiena.com