hygiena KIT230032 Magnetic Preparation Kit III User Guide

: June 17, 2024
: Hygiena

Table of Contents

hygiena KIT230032 Magnetic Preparation Kit III
Product Information
Product Usage Instructions
PROGRAM SETUP
DATA INTERPRETATION
PREPARATION OF THE PCR MIX
References
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hygiena KIT230032 Magnetic Preparation Kit III

Product Information

Specifications

Product Name: Animal Detection 2 LyoKit
Product No.: KIT230032 (LP), KIT230033 (RP)
Revision: A, November 2023
PCR kit for the qualitative detection of chicken (Gallus gallus), turkey (Meleagris gallopavo), and poultry DNA
Detection of the whole class of Aves, including Goose (Anser), duck (Anas), ostrich (Struthio), guinea fowl (Numida), and other bird DNA

Program Setup
Before setting up the PCR reactions, program your real-time PCR instrument by selecting the following channels:

FAM (chicken)
HEX (turkey)
ROX (poultry/Aves)
Cy5 (Internal Control)

Follow the cycling conditions as specified in the manual.

Data Interpretation
Verify results of positive and negative controls before interpreting sample results. Always compare samples to positive and negative controls.

Preparation of the PCR Mix

Place strips in the rack after taking them out of the aluminium bag.
Decap the strips immediately before filling.
Add samples and controls.
Seal the tubes with the provided cap strips.
Mix thoroughly to resuspend the pellet.
Centrifuge briefly to settle the contents.
Start the real-time PCR run as per instructions.

Product Usage Instructions

Place Strips in Rack: Take PCR tube strips out of the bag and place them in a suitable rack.
Decap: Open strips just before filling and discard caps immediately.
Add Samples and Controls: Add the required samples and controls to the tubes.
Seal: Carefully seal the tubes with the provided cap strips.
Mix: Thoroughly mix to resuspend the pellet by either gentle tapping or pipetting up and down.
Centrifuge: Briefly spins the strips in a centrifuge to settle the contents.
Start Real-Time PCR Run: Cycle the samples following the specified conditions. Place tubes in a balanced order in the cycler.

FAQ

Q: What is the purpose of the Animal Detection 2 LyoKit?
A: The kit is designed for the qualitative detection of various bird DNA including chicken, turkey, and poultry for food testing purposes.
Q: Can I reuse the PCR tube strips?
A: It is recommended to use fresh PCR tube strips for each new set of reactions to prevent contamination.

Product No.

KIT230032 (LP)
KIT230033 (RP)

PCR kit for the qualitative detection of chicken (Gallus gallus), turkey (Meleagris gallopavo) and poultry DNA [detection of the whole class of Aves, including goose (Anser), duck (Anas), ostrich (Struthio), chicken (Gallus), turkey (Meleagris), guinea fowl (Numida) and other bird DNA].

PROGRAM SETUP

Program your real-time PCR instrument before setting up the PCR reactions. Select the following channels:

FAM (chicken), HEX (turkey), ROX (poultry / Aves) and Cy5 (Internal Control). $hygiena-KIT230032-Magnetic-Preparation-Kit-III-fig- $1$$
Pre-incubation: 1 cycle
- Step 1: 37 °C for 4 min
- Step 2: 95 °C for 5 min
Ampliﬁ cation: 35 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 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).

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	HEX	ROX	Cy5	Result Interpretation
+	+	+	+ or –	Positive for chicken, turkey, poultry
+	–	+	+ or –	Positive for chicken, poultry
–	+	+	+ or –	Positive for turkey, poultry
–	–	+	+ or –	Positive for poultry, negative for chicken and turkey
–	–	–	+	Negative for chicken, turkey, poultry
–	–	–	–	Invalid

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 (from Sample Preparation Kit III), Negative Control (colourless cap) or Control Template (purple cap) into respective wells.
- For samples prepared with StarPrep Five, use 22 µL PCR-grade water + 3 µL sample for a total volume of 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.