MAN0009929 RapidFinderPoultry UG
MAN0009929 RapidFinderPoultry UG
MAN0009929 RapidFinderPoultry UG
USER GUIDE
Trademarks: All trademarks are the property of Thermo Fisher Scientific and its subsidiaries unless otherwise specified. TaqMan is a
registered trademark of Roche Molecular Systems, Inc., used under permission and license. Imegen agro is a trademark of Instituto de
Medicina Genómica SL.
©2021 Thermo Fisher Scientific Inc. All rights reserved.
Contents
Product description . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
Kit contents and storage . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
Materials required but not provided . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6
■ CHAPTER 2 Methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
■ APPENDIX A Troubleshooting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11
■ APPENDIX D Safety . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 16
Chemical safety . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17
Biological hazard safety . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 18
Product description
Identification of meat species present in food samples is an essential step for verification of origin
and traceability of raw materials, as well as for quality control of handling and cleaning processes in
production lines. The Thermo Scientific™ RapidFinder™ Poultry ID Kit enables real-time PCR detection
of poultry DNA that is present in food and feed samples. The kit detects blends containing ≥0.01% of
poultry DNA.
The kit includes:
• All reagents necessary for the real‐time PCR reaction—specific FAM™-labeled probe and primers
for poultry mitochondrial DNA, DNA polymerase enzyme, and other buffer components.
• An internal positive control (IPC)—VIC™-labeled probe, primers, and template, to monitor for PCR
inhibition.
• A Positive Control, to confirm poultry DNA detection.
Item Source
Applied Biosystems™ QuantStudio™ 5 Real‑Time PCR System Contact your local microbiology
RapidFinder™ Analysis Software v1.2 or later sales representative.
Equipment
Adjustable micropipettors (10 µL, 20 µL, 200 µL) Available through the Thermo
Fisher Microbiology ordering
Benchtop microcentrifuge with adaptors for PCR plates and/or tubes process. See thermofisher.com/
plastics for more information.
Laboratory mixer (Vortex mixer or equivalent)
Optical reaction plates and covers, or optical PCR tubes and caps
Reagents
3. Add 5 µL of DNA sample (10 ng/µL), mock-purified sample (negative extraction control),
nuclease-free water (no-template control), or Positive Control to the appropriate wells.
4. Seal each plate or tube, mix, then centrifuge briefly to bring the contents to the bottom.
Stage 1 Stage 2
Setting
Enzyme activation PCR
Number of cycles 1 (Hold) 36
Denature Anneal/extend[1]
Temperature 95°C 95°C 60°C
Time 10 minutes 15 seconds 1 minute
[1] Fluorescence is acquired during the annealing/extension stage.
2. Load the reactions, run the thermal cycler program and collect real-time amplification data.
Analyze results
The general process for analyzing results is described in this section. The details of data analysis
depend on the real-time PCR instrument that you use; see the appropriate user guide for instructions
on how to analyze your data.
1. View the amplification plots for all reactions to make sure that they appear normal.
4. Check that the results obtained in all control wells are as expected. For unexpected control results,
see Appendix A, “Troubleshooting”.
5. Establish the positive cut-off value for the test samples and assign results:
Ct (cut-off) = Ct (Positive Control) + 3.32
Interpretation of results
Interpret unknown sample results according to the following table.
Quantitative analysis
For samples with positive results, the percentage of poultry DNA with respect to total animal DNA
can be quantified using the RapidFinder™ Poultry ID Kit in combination with the RapidFinder™ Quant
Multi-Meat Set (Cat. No. A24399).
The RapidFinder™ Quant Multi-Meat Set includes the Multi-Meat Standard, a plasmid DNA quantitation
standard containing individual species-specific DNA targets and a highly conserved animal-specific
mitochondrial genomic region (total animal DNA target). The kit also includes a TaqMan™ assay for the
total animal DNA target. Both species-specific and total animal DNA present in each sample can be
quantified relative to the Multi-Meat Standard using primer and probe sets from both kits. For detailed
instructions, see the RapidFinder™ Quant Multi-Meat Set User Guide (Pub. No. MAN0009930).
Note: The Positive Control included in the RapidFinder™ Poultry ID Kit is not intended for use as a
quantitation standard.
Note: The primers and probes in this kit have been designed to detect any kind of bird species. It is
more common to detect poultry meat in food samples, but it is possible to use this kit to detect bird
species DNA in general.
Chicken Detected
Turkey Detected
Duck Detected
Ostrich Detected
Goose Detected
To avoid amplicon contamination of samples, follow these guidelines when preparing or handling
samples for PCR amplification:
• Wear clean gloves and a clean lab coat (not previously worn while handling amplified products or
used during sample preparation).
• Change gloves whenever you suspect that they are contaminated.
• Maintain separate areas and dedicated equipment and supplies for:
– Sample preparation and reaction setup.
– Amplification and analysis of products.
• Do not bring amplified products into the reaction setup area.
• Open and close all sample tubes carefully. Avoid splashing or spraying samples.
• Keep reactions and components capped as much as possible.
• Use a positive-displacement pipettor or aerosol-resistant barrier pipette tips.
• Do not open reaction tubes after PCR.
• Do not autoclave reaction tubes after PCR.
• Clean lab benches and equipment periodically with 10% bleach solution or DNAZap™ Solutions
(Cat. No. AM9890). After cleaning with bleach we recommend a rinse with an ethanol solution
because bleach will rust stainless steel.
WARNING! GENERAL SAFETY. Using this product in a manner not specified in the user
documentation may result in personal injury or damage to the instrument or device. Ensure that
anyone using this product has received instructions in general safety practices for laboratories and
the safety information provided in this document.
· Before using an instrument or device, read and understand the safety information provided in the
user documentation provided by the manufacturer of the instrument or device.
· Before handling chemicals, read and understand all applicable Safety Data Sheets (SDSs) and use
appropriate personal protective equipment (gloves, gowns, eye protection, and so on). To obtain
SDSs, see the “Documentation and Support” section in this document.
Chemical safety
WARNING! GENERAL CHEMICAL HANDLING. To minimize hazards, ensure laboratory personnel
read and practice the general safety guidelines for chemical usage, storage, and waste provided
below. Consult the relevant SDS for specific precautions and instructions:
· Read and understand the Safety Data Sheets (SDSs) provided by the chemical manufacturer
before you store, handle, or work with any chemicals or hazardous materials. To obtain SDSs, see
the "Documentation and Support" section in this document.
· Minimize contact with chemicals. Wear appropriate personal protective equipment when handling
chemicals (for example, safety glasses, gloves, or protective clothing).
· Minimize the inhalation of chemicals. Do not leave chemical containers open. Use only with
sufficient ventilation (for example, fume hood).
· Check regularly for chemical leaks or spills. If a leak or spill occurs, follow the manufacturer
cleanup procedures as recommended in the SDS.
· Handle chemical wastes in a fume hood.
· Ensure use of primary and secondary waste containers. (A primary waste container holds the
immediate waste. A secondary container contains spills or leaks from the primary container.
Both containers must be compatible with the waste material and meet federal, state, and local
requirements for container storage.)
· After emptying a waste container, seal it with the cap provided.
· Characterize (by analysis if needed) the waste generated by the particular applications, reagents,
and substrates used in your laboratory.
· Ensure that the waste is stored, transferred, transported, and disposed of according to all local,
state/provincial, and/or national regulations.
· IMPORTANT! Radioactive or biohazardous materials may require special handling, and disposal
limitations may apply.
· Lire et comprendre les fiches de données de sécurité (FDS) fournies par le fabricant avant de
stocker, de manipuler ou d’utiliser les matériaux dangereux ou les produits chimiques. Pour obtenir
les FDS, se reporter à la section « Documentation et support » du présent document.
· Limiter les contacts avec les produits chimiques. Porter des équipements de protection appropriés
lors de la manipulation des produits chimiques (par exemple : lunettes de sûreté, gants ou vête‐
ments de protection).
· Limiter l’inhalation des produits chimiques. Ne pas laisser les récipients de produits chimiques
ouverts. Ils ne doivent être utilisés qu’avec une ventilation adéquate (par exemple, sorbonne).
· Vérifier régulièrement l’absence de fuite ou d’écoulement des produits chimiques. En cas de fuite
ou d’écoulement d’un produit, respecter les directives de nettoyage du fabricant recommandées
dans la FDS.
· Manipuler les déchets chimiques dans une sorbonne.
· Veiller à utiliser des récipients à déchets primaire et secondaire. (Le récipient primaire contient les
déchets immédiats, le récipient secondaire contient les fuites et les écoulements du récipient pri‐
maire. Les deux récipients doivent être compatibles avec les matériaux mis au rebut et conformes
aux exigences locales, nationales et communautaires en matière de confinement des récipients.)
· Une fois le récipient à déchets vidé, il doit être refermé hermétiquement avec le couvercle fourni.
· Caractériser (par une analyse si nécessaire) les déchets générés par les applications, les réactifs et
les substrats particuliers utilisés dans le laboratoire.
· Vérifier que les déchets sont convenablement stockés, transférés, transportés et éliminés en res‐
pectant toutes les réglementations locales, nationales et/ou communautaires en vigueur.
· IMPORTANT ! Les matériaux représentant un danger biologique ou radioactif exigent parfois une
manipulation spéciale, et des limitations peuvent s’appliquer à leur élimination.
WARNING! 4L Reagent and Waste Bottle Safety. Four-liter reagent and waste bottles can crack
and leak. Each 4-liter bottle should be secured in a low-density polyethylene safety container with the
cover fastened and the handles locked in the upright position.
WARNING! BIOHAZARD. Biological samples such as tissues, body fluids, infectious agents,
and blood of humans and other animals have the potential to transmit infectious diseases.
Conduct all work in properly equipped facilities with the appropriate safety equipment (for example,
physical containment devices). Safety equipment can also include items for personal protection,
such as gloves, coats, gowns, shoe covers, boots, respirators, face shields, safety glasses, or
goggles. Individuals should be trained according to applicable regulatory and company/ institution
requirements before working with potentially biohazardous materials. Follow all applicable local,
state/provincial, and/or national regulations. The following references provide general guidelines when
handling biological samples in laboratory environment.
· U.S. Department of Health and Human Services, Biosafety in Microbiological and Biomedical
Laboratories (BMBL), 6th Edition, HHS Publication No. (CDC) 300859, Revised June 2020
https://www.cdc.gov/labs/pdf/CDC-BiosafetymicrobiologicalBiomedicalLaboratories-2020-
P.pdf
· Laboratory biosafety manual, fourth edition. Geneva: World Health Organization; 2020 (Laboratory
biosafety manual, fourth edition and associated monographs)
www.who.int/publications/i/item/9789240011311
Phone: Visit thermofisher.com/support, select the link for phone support, then select the appropriate
country from the dropdown list.
Note: For SDSs for reagents and chemicals from other manufacturers, contact the
manufacturer.
Related documentation
Document Publication number
PCR Starter Kit for 96-well blocks, 0.2 mL, User Guide A24829
8 July 2021