hygiena KIT230076 Beer Screening 2 LyoKit User Guide

: May 15, 2024
: Hygiena

Table of Contents

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

hygiena KIT230076 Beer Screening 2 LyoKit

hygiena KIT230076 Beer Screening 2 LyoKit

Important Information

PCR kit for screening of >30 beer spoilage bacteria with detection of the genera Lactobacillus / Pediococcus and Pectinatus / Megasphaera in two separate channels plus detection of hop-tolerance genes horA and horC. 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 (beer-spoiling Lactobacillus and Pediococcus), HEX (beer-spoiling Pectinatus and Megasphaera), 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 (H2O) 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	ROX	ATTO 490LS	Result Interpretation
+	+	v	+ or –	Positive for beer-spoilage bacteria and hop-tolerance

related genes
+| +| –| + or –| Positive for Lactobacillus and/or Pediococcus and Pectinatus and/or Megasphaera
+| –| +| + or –| Positive for Lactobacillus and/or Pediococcus and hop- tolerance related genes
+| –| –| + or –| Positive for Lactobacillus and/or Pediococcus
–| +| –| + or –| Positive for Pectinatus and/or Megasphaera
–| –| +| + or –| Positive for hop-tolerance related genes
–| –| –| +| Negative for beer-spoilage bacteria, hop-tolerance related genes
–| –| –| –| 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, Negative Control (colorless cap) or Control Template (purple cap) into respective wells.
If using less volume, add PCR-grade H2O 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 first and last column.