hygiena KIT230073 Beer Screening 1 LyoKit User Guide

: May 15, 2024
: Hygiena

Table of Contents

IMPORTANT INMFORMATION
PROGRAM SETUP
DATA INTERPRETATION
PREPARATION OF THE PCR MIX
Customer Support
References
Read User Manual Online (PDF format)
Download This Manual (PDF format)

hygiena KIT230073 Beer Screening 1 LyoKit

hygiena KIT230073 Beer Screening 1 LyoKit

IMPORTANT INMFORMATION

Product No. KIT230073 (DP)

PCR kit for screening of >30 beer spoilage bacteria of the genera Lactobacillus, Pediococcus, Pectinatus and Megasphaera, including identification of L. brevis and detection of hop-tolerance genes horA and horC on a real-time PCR cycler.
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 dyes:

FAM (beer-spoilage bacteria), HEX (Lactobacillus brevis), ROX (hop-tolerance related genes) and ATTO 490LS (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 10 sec
*Step 2:** 60 °C for 60 sec
Fluorescence detection

For the Dualo 32® Beverage real-time PCR instrument, please open the software, click on ‘New’, and select the respective template file. Template files can be added by clicking on ‘Add’ in the ‘Select template file’ window.

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 dye and interpret results as described in the table.

FAM	HEX	ROX	ATT0490LS	Result Interpretation
+	+	+	+ or –	Positivefor beer-spoilage bacteria, L brevis,* hop-

tolerance related genes
+| –| +| + or –| Positive for beer-spoilage bacteria, hop-tolerance related genes
+| +| –| + or –| Positive for beer-spoilage bacteria, L. brevis*
+| –| –| + or –| Positive for beer-spoilage bacteria
–| –| +| + or –| Positive for hop-tolerance related genes
–| –| –| +| Negative for beer-spoilage bacteria, hop-tolerance related genes
–| –| –| –| Invalid

*including Lactobacillus sp. DSM 6265 (“L. brevisimilis”)

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, Negative Control (colorless cap) or Control Template (purple cap) into respective wells. If using less volume, add PCR-grade H2 O to reach 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 fi rst and last column.