Hygiena foodproof KIT230127 User manual

RR-KIT230127-28-RevA
foodproof®is a registered trademark of Hygiena. The above mentioned real-time PCR instruments are registered trademarks of their respective holders.
Pre-incubation: 1 cycle
Step 1: 37 °C for 4 min
Step 2: 95 °C for 5 min
Amplification: 35 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 : 60 °C for 50 sec
Step 3**: ramp up to 80 °C
1 cycle 1 cycle
4 min 5 min 5 sec | 60 sec*50 sec, 50 sec, ramp**
35 cycles
*Fluorescence detection ** Fluorescence detection during 60 - 80 °C ramp with 1 measurement/°C
95 °C
80 °C
60 °C
37 °C
Animal Detection 1 LyoKit
Ready Reference Guide
PCR kit for the qualitative detection of porcine, bovine and equine with differentiation of horse, donkey and zebra.
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:
uFAM (porcine), HEX (bovine), ROX (equine: horse, donkey, zebra) and
Cy5 (Internal Control).
For CFX96TM real time PCR cycler step 2 of melting curve is: 50 °C for 50 seconds. For ABI 7500 Fast, step 2
of melting curve is: 50 °C for 60 seconds, step 3: ramp up to 85 °C. 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 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).
Product No. KIT230127 (LP), KIT230128 (RP)
Revision A, November 2023
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.
foodproof®
Channel Result Interpretation - Melting Curve
ROX Donkey: 64 - 67 °C Zebra: 68 - 70.5 °C Horse: 71 - 74 °C
FAM HEX ROX Cy5 Result Interpretation - Amplification
+ + + + or - Positive for porcine, bovine and equine
- + + + or - Positive for bovine and equine
+ - + + or - Positive for porcine and equine
+ + - + or - Positive for porcine and bovine
- + - + or - Positive for bovine
+ - - + or - Positive for porcine
- - + + or - Positive for equine
- - - + Negative for porcine, bovine and equine
----Invalid
The Control Template contains only donkey and horse. Therefore, two peaks will be displayed. Please use the
Control Template as a reference. Melting curve peaks can vary about ± 1 °C depending on PCR instrument used.

foodproof®
Animal Detection 1 LyoKit
KIT230127 / 28
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
3. ADD SAMPLES AND CONTROLS
Pipette 25 µL of samples (from Sample Preparation Kit III), Negative Con-
trol (colorless 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.
2. DECAP
Immediately before filling, carefully open strips and discard caps.
Do not leave open longer than necessary.
1. 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.
4. SEAL
Carefully seal the tubes with the provided 8-cap strips.
6. CENTRIFUGE
Briefly spin strips, e.g., 5 seconds at 500 - 1,000 x g, in a suitable
centrifuge.
5. MIX
Resuspend pellet after sealing by mixing thoroughly.
Alternatively, resuspend pellet by pipetting up and down
multiple times in Step 3.
7. 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.
PREPARATION OF THE PCR MIX
Take appropriate precautions to prevent contamination, e.g., by using filter tips and wearing gloves.