hygiena KIT230049 foodproof Salmonella Detection Kit User Guide

: June 17, 2024
: Hygiena

Table of Contents

hygiena KIT230049 foodproof Salmonella Detection Kit
Specifications
Product Usage Instructions
FAQ
PROGRAM SETUP
DATA INTERPRETATION
PREPARATION OF THE PCR MIX
ABOUT COMPANY
References
Read User Manual Online (PDF format)
Download This Manual (PDF format)

hygiena KIT230049 foodproof Salmonella Detection Kit

Specifications

Product Name: Salmonella Detection Kit
Product No.: KIT230049, KIT230050
Revision: A, December 2023
PCR kit for the qualitative detection of Salmonella DNA using real-time PCR instruments
Contains probes with TAMRA as quencher and no passive reference dye

Product Usage Instructions

Program Setup:
Before setting up the PCR reactions, program your real-time PCR instrument with the following settings:

Select channels: FAM (Salmonella) and VIC (Internal Control)
Run a program of 1 cycle with timings – 4 min | 5 min, followed by 50 cycles of 5 sec | 60 sec* with fluorescence detection

Data Interpretation
Verify results of positive (Control Template) and negative controls (H2O) before interpreting sample results. Always compare samples to positive and negative controls. Interpret results based on the table provided.

Preparation of the PCR Mix:

Take appropriate precautions to prevent contamination, e.g., use filter tips and wear gloves.
Thaw reagents, mix (do not vortex!) and briefly spin vials before opening.
Prepare PCR Mix: Mix carefully but thoroughly by pipetting up and down.
Add PCR Mix:
Add Samples and Controls:
Seal: Seal strips/plate accurately.
Centrifuge: Briefly spin strips/plate in a suitable centrifuge.
Start Real-Time PCR Run: Cycle samples as described in the program setup.
For food testing purposes. FOR IN VITRO USE ONLY. Made in Germany.

FAQ

Can this kit be used for detecting other pathogens?
No, this kit is specifically designed for the qualitative detection of Salmonella DNA only.
Is it necessary to wear gloves during the entire process?
It is strongly recommended to wear gloves to prevent contamination, especially when handling samples and reagents.

PROGRAM SETUP

Program your real-time PCR instrument before setting up the PCR reactions. Select the following channels:
FAM (Salmonella) and VIC (Internal Control).
As an alternative to VIC, HEX can be used. For the PikoReal® 24, Yakima Yellow has to be selected.

hygiena-KIT230049-foodproof-Salmonella-Detection-Kit-
fig-2

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
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.

hygiena-KIT230049-foodproof-Salmonella-Detection-Kit-
fig-3

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.