hygiena KIT230018 Yersinia Enterocolitica Plus Pseudotuberculosis Detection Kit User Guide
- June 1, 2024
- Hygiena
Table of Contents
hygiena KIT230018 Yersinia Enterocolitica Plus Pseudotuberculosis
Detection Kit
Product Information
Specifications:
- Product Name: Yersinia enterocolitica plus pseudotuberculosis Detection Kit
- Revision: A, December 2023
- Product No.: KIT230018
- PCR kit for the qualitative detection of Yersinia enterocolitica plus Yersinia pseudotuberculosis DNA using real-time PCR instruments
Product Usage Instructions
Program Setup:
Program your real-time PCR instrument before setting up the PCR reactions.
Select the following channels: FAM (Yersinia enterocolitica), HEX (Yersinia
pseudotuberculosis), and ROX
(Internal Control).
Data Interpretation:
Verify results of positive (Control Template) and negative controls (H2O)
before interpreting sample results. Always compare samples to positive and
negative controls. Review data from each channel and interpret results as
described in the table provided.
Preparation of the PCR Mix:
- 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 section.
FAQ
-
Q: Can this kit be used for food testing purposes?
A: Yes, this kit can be used for food testing purposes. -
Q: How many reactions can be performed with this kit?
A: This kit allows for 64 reactions to be performed. -
Q: Is this kit for in vitro use only?
A: Yes, this detection kit is for in vitro use only.
PCR kit for the qualitative detection of Yersinia enterocolitica plus Yersinia pseudotuberculosis 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 (Yersinia enterocolitica), HEX (Yersinia pseudotuberculosis) and ROX
(Internal Control).
Pre-incubation: 1 cycle
Step 1: 37 °C for 4 min
Step 2: 95 °C for 5 min
Amplification: 50 cycles
Step 1 : 95 °C for 5 s
*Step 2:** 60 °C for 60 s
- Fluorescence detection
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 5 (Product No.: KIT230011).
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 | Result Interpretation |
|---|---|---|---|
| + | + | + or – | Positive for Yersinia enterocolitica plus _Yersinia |
pseudotuberculosis_
–| +| + or –| Positive for Yersinia pseudotuberculosis
+| –| + or –| Positive for Yersinia enterocolitica
–| –| +| Negative for Yersinia enterocolitica plus Yersinia
pseudotuberculosis
–| –| –| Invalid
foodproof® is a registered trademark of Hygiena. The above mentioned real-time PCR instruments are registered trademarks of their respective holders.
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
Briefly spin strips/plate in a suitable centrifuge.
START REAL-TIME PCR RUN
Cycle samples as described above.
foodproof® Yersinia entero-colitica plus pseudotubercu-losis Detection Kit
KIT230018
Kit for 64 reactions Store kit at -15 to -25 °C
For food testing purposes FOR IN VITRO USE ONLY Made in Germany
Hygiena® | Camarillo, CA 93012 USA
Email:
[email protected]
Website: www.hygiena.com