Product Overview
This document provides a ready reference for the Hygiena foodproof® Yeast and Mold Quantification LyoKit. It is designed for the quantitative detection of yeast and mold using real-time PCR instruments.
Product Numbers: KIT230112 (LP), KIT230113 (RP), KIT230114 (DP)
For detailed information, users are strongly recommended to consult the complete product manual available on the Hygiena website.
Program Setup
Before preparing PCR reactions, program your real-time PCR instrument. The kit utilizes the following channels:
- FAM (for yeast and mold detection)
- HEX (for Internal Control)
The recommended PCR cycling conditions are detailed, including pre-incubation and amplification steps with specific temperatures and durations.
Note: This kit uses probes with a non-fluorescent "dark" quencher and does not require a passive reference dye.
Preparation of Standard Curve
To prepare the standard curve, use the Quantification Standard (vial 2, purple cap) and Negative Control (vial 3, colorless cap). Dilutions are prepared by pipetting 90 µL of Negative Control into a new tube for each step, then transferring 10 µL from the preceding step. Mixing is crucial between steps.
A typical experiment includes 9 wells for standards (duplicates) and negative control, plus additional wells for food samples.
| Dilution Step | Dilution Factor | Concentration to Be Entered as Standard (GE/reaction) |
|---|---|---|
| 1 | Undiluted | 10,000 |
| 2 | 1:10 | 1,000 |
| 3 | 1:100 | 100 |
| 4 | 1:1,000 | 10 |
Preparation of the PCR Mix
Follow these steps to prepare the PCR mix, taking precautions against contamination:
- Place Strips in Rack: Remove PCR tube strips from the aluminum bag and place them in a suitable rack.
- Decap: Open strips carefully just before filling and discard caps.
- Add Samples and Controls: Pipet 25 µL of samples, standards, and Negative Control into respective wells. Adjust volume with Negative Control if sample volume is less than 25 µL.
- Seal: Seal the tubes securely with the provided 8-cap strips.
- Mix: Resuspend the pellet by thorough mixing.
- Centrifuge: Briefly spin the strips (e.g., 5 seconds at 500-1,000 x g).
- Start Real-Time PCR Run: Place tubes in the cycler in a balanced order.
Product Information
Kit: foodproof® Yeast and Mold Quantification LyoKit
Contents: Kit for 96 reactions
Storage: Store kit at 2 to 8 °C
Application: For food testing purposes
Usage: FOR IN VITRO USE ONLY
Manufacturer: Hygiena, Camarillo, CA 93012 USA
Contact: diagnostics.support@hygiena.com | www.hygiena.com
Innovative Diagnostic Solutions for Hygiene & Food Safety | Hygiena
