hygiena KIT230115 LP Porcine Detection LyoKit User Guide
- June 16, 2024
- Hygiena
Table of Contents
foodproof®
Porcine Detection LyoKit
Ready Reference Guide
Revision A, November 2023
PCR kit for the qualitative detection of Sus scrofa DNA using real-time PCR
instruments.
Before starting, it is strongly recommended to read the entire product manual
available on our website.
Product No. KIT230115 (LP), KIT230116 (RP)
PROGRAM SETUP
Program your real-time PCR instrument before setting up the PCR reactions.
Select the following channels:
▶ FAM (Sus scrofa DNA) and HEX (Internal Control).
Pre-incubation: 1 cycle
Step 1: 37 °C for 4 min
Step 2: 95 °C for 10 min
Amplification: 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 use of a passive reference dye must be specified. This kit contains probes with a non- fluorescent “dark” quencher and no passive reference dye.
DATA INTERPRETATION
Verify results of positive (Control Template) and negative (H² O) 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 | Result Interpretation |
|---|---|---|
| + | + or + | Positive for Sus scrofa DNA |
| – | + | Negative for Sus scrofa DNA |
| – | – | Invalid |
PREPARATION OF THE PCR MIX
Take appropriate precautions to prevent contamination, e.g., by using filter 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 filling, carefully open strips and discard caps.
Do not leave open longer than necessary. -
ADD SAMPLES AND CONTROLS
Pipette 25 µL of samples (from Sample Preparation Kit III), Negative Control (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. -
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
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.
foodproof®
Porcine Detection LyoKit
KIT230115 / 16
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
Documents / Resources
|
hygiena KIT230115 LP Porcine Detection
LyoKit
[pdf] User Guide
KIT230115 LP Porcine Detection LyoKit, KIT230115 LP, Porcine Detection LyoKit,
Detection LyoKit, LyoKit
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