Thermo Fisher Scientific VetMAX AIV Reagents Mode d'emploi

Marque: Thermo Fisher Scientific

Modèle: VetMAX AIV Reagents

Catégorie: Kits de voiture

Taper: Mode d'emploi

For Research Use Only. Not for use in diagnostic procedures.

VetMAX™ AIV Reagents

Catalog Numbers 4405543

Publication Number MAN0018920

Revision A.0

Life Technologies Corporation | 2130 Woodward Street | Austin, TX 78744

For descriptions of symbols on product labels or product documents, go to thermoﬁsher.com/symbols-deﬁnition.

The information in this guide is subject to change without notice.

DISCLAIMER: TO THE EXTENT ALLOWED BY LAW, THERMO FISHER SCIENTIFIC INC. AND/OR ITS AFFILIATE(S) WILL NOT BE LIABLE FOR SPECIAL,

INCIDENTAL, INDIRECT, PUNITIVE, MULTIPLE, OR CONSEQUENTIAL DAMAGES IN CONNECTION WITH OR ARISING FROM THIS DOCUMENT,

INCLUDING YOUR USE OF IT.

Revision history: Pub. No. MAN0018920

Revision Date Description

A.0 29 October 2019

Converted the existing manual (4412867B, dated 30Jun2011) to the

current document template, with associated updates to the limited

license information, warranty, trademarks, and logos. Edited to remove

the Detection Enhancer as an included kit component.

Important Licensing Information: These products may be covered by one or more Limited Use Label Licenses. By use of these products, you accept

the terms and conditions of all applicable Limited Use Label Licenses.

TRADEMARKS: All trademarks are the property of Thermo Fisher Scientiﬁc and its subsidiaries unless otherwise speciﬁed.

Contents

■VetMAX™ AIV Reagents .................................................... 4

Product description ............................................................. 4

Contents and storage ............................................................ 4

Required materials not supplied .................................................. 4

Guidelines for input RNA ......................................................... 5

Set up, then run the real-time RT-PCR reaction ..................................... 5

Supplemental information ........................................................ 6

Good laboratory practices for PCR and RT-PCR ................................. 6

Customer and technical support .................................................. 6

Limited product warranty ........................................................ 7

■Safety ....................................................................... 8

Chemical safety ................................................................. 9

VetMAX

™

AIV Reagents User Guide

VetMAX™ AIV Reagents

WARNING! Read the Safety Data Sheets (SDSs) and follow the handling

instructions. Wear appropriate protective eyewear, clothing, and gloves. Safety

Data Sheets (SDSs) are available from thermosher.com/support.

Product description

TaqMan™ AIV-Matrix Reagents enable one-step, real-time RT-PCR targeting the Avian

Inuenza Virus matrix gene RNA and Xeno™ RNA Control. The RT-PCR tolerates a

wide range of input RNA amounts, but it is not recommended to use more than 100

ng of RNA.

Contents and storage

Component Amount[1] Storage[2]

2X RT-PCR Buffer 1.375 mL

–20°C

25X RT-PCR Enzyme Mix 110 µL

25X AIV-Matrix Primer

Probe Mix 110 µL

Nuclease-free Water 1.75 mL —

[1] Reagents for 100 25-µL RT-PCRs are supplied.

[2] Store the reagents in a non-frost-free freezer.

Required materials not supplied

Unless otherwise indicated, all materials are available through thermosher.com.

MLS: Fisher Scientic (sherscientic.com) or other major laboratory supplier.

Item Source

(Optional)

Detection Enhancer

Detection Enhancer ‑ 100 rxn A44810

Detection Enhancer ‑ 500 rxn A44941

Detection Enhancer ‑ 1000 rxn A44811

VetMAX

™

AIV Reagents User Guide

Guidelines for input RNA

• Add Xeno™ RNA Control to the lysis solution used for RNA isolation, to serve as

a positive control both for recovery of RNA and for the RT-PCR. Add 5,000–

10,000 copies of Xeno™ RNA Control per isolation (0.5–1.0 µL at 10,000

copies/µL).

• When isolating RNA from cell-free sample sources such as serum, add carrier

RNA to the lysis solution to maximize RNA recovery. Include carrier RNA even if

you have added Xeno™ RNA Control to the lysis solution.

• It is important to use RNA that is free of RT-PCR inhibitors in the procedure.

• We recommend using a MagMAX™ RNA isolation kit that is appropriate for your

sample type. Go to www.thermosher.com, then search for

MagMAX

• For cell-free sample types, such as serum, we recommend using the MagMAX™

CORE Nucleic Acid Purication Kit (Cat. No. A32700 or A32702).

Set up, then run the real-time RT-PCR reaction

• Calculate the number of required reactions. Scale reaction components based on

the single-reaction volumes, then include 5–10% overage, unless otherwise

indicated.

• Include duplicate no template controls (NTCs) using nuclease-free water in place

of sample.

1. Program the real-time PCR instrument according to the manufacturer's

instructions.

• Passive reference dye: ROX™ (included in the RT-PCR Buer)

• Reaction volume: 25 µL

Target Reporter Quencher

AIV-Matrix FAM™ Dye BHQ™-1 Dye

Xeno™ RNA Control CF560 Dye [1]

[1] CAL Fluor™ Orange 560 (CF560) dye has an absorbance maximum of 538 nm and an emission

maximum of 559 nm. When using an Applied Biosystems™ Real-Time PCR System, it is best to calibrate

with CF560 dye. If not available, use the VIC™ dye detector for CF560 dye.

2. Prepare each RT-PCR master mix on ice.

Component Amount

RT-PCR master mix

2X RT-PCR Buffer 12.5 µL

25X AIV-Matrix Primer Probe Mix 1 µL

25X RT-PCR Enzyme Mix 1 µL

(Optional)

Detection Enhancer (1.67 µL)

Total volume of master mix per

reaction

14.5 µL

(16.17 µL including

optional Detection

Enhancer)

VetMAX™ AIV Reagents

Guidelines for input RNA

VetMAX

™

AIV Reagents User Guide

3. Distribute RT-PCR master mix to the wells of a PCR plate or to PCR tubes.

4. Add sample or controls to the wells or tubes containing RT-PCR master mix

(25 µL nal volume per reaction).

5. Run the thermal cycler program.

Note: Refer to AgPath-ID™ One-Step RT-PCR Reagents for an example of a

thermal cycler program.

Supplemental information

• Wear clean gloves and a clean lab coat.

– Do not wear the same gloves and lab coat that you have previously used

when handling amplied products or preparing samples.

• Change gloves if you suspect that they are contaminated.

• Maintain separate areas and dedicated equipment and supplies for:

– Sample preparation and reaction setup.

–Amplication and analysis of products.

• Do not bring amplied 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 pipeor or aerosol-resistant barrier pipee tips.

• Clean lab benches and equipment periodically with 10% bleach solution or DNA

decontamination solution.

Customer and technical support

Visit thermosher.com/support for the latest service and support information.

• Worldwide contact telephone numbers

• Product support information

– Product FAQs

– Software, patches, and updates

– Training for many applications and instruments

• Order and web support

• Product documentation

– User guides, manuals, and protocols

–Certicates of Analysis

– Safety Data Sheets (SDSs; also known as MSDSs)

Note: For SDSs for reagents and chemicals from other manufacturers,

contact the manufacturer.

Good laboratory

practices for PCR

and RT-PCR

VetMAX™ AIV Reagents

Supplemental information

VetMAX

™

AIV Reagents User Guide

Limited product warranty

Life Technologies Corporation and/or its aliate(s) warrant their products as set forth

in the Life Technologies' General Terms and Conditions of Sale at

www.thermosher.com/us/en/home/global/terms-and-conditions.html. If you have

any questions, please contact Life Technologies at www.thermosher.com/support.

VetMAX™ AIV Reagents

Limited product warranty

VetMAX

™

AIV Reagents User Guide

Safety

WARNING! GENERAL SAFETY. Using this product in a manner not specied

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.

VetMAX

™

AIV Reagents User Guide

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 specic 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 sucient 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.

AVERTISSEMENT ! PRÉCAUTIONS GÉNÉRALES EN CAS DE

MANIPULATION DE PRODUITS CHIMIQUES. Pour minimiser les risques,

veiller à ce que le personnel du laboratoire lise aentivement et mee en œuvre

les consignes de sécurité générales relatives à l’utilisation et au stockage des

produits chimiques et à la gestion des déchets qui en découlent, décrites ci-

dessous. Consulter également la FDS appropriée pour connaître les précautions

et instructions particulières à respecter :

·Lire et comprendre les ches 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 : lunees de sûreté, gants ou vêtements 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).

Safety

Chemical safety

VetMAX

™

AIV Reagents User Guide

·Vérier 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 neoyage 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 primaire. Les deux récipients doivent

être compatibles avec les matériaux mis au rebut et conformes aux exigences

locales, nationales et communautaires en matière de connement 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.

·Vérier que les déchets sont convenablement stockés, transférés, transportés

et éliminés en respectant 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.

Safety

Chemical safety

VetMAX

™

AIV Reagents User Guide

thermoﬁsher.com/support | thermoﬁsher.com/askaquestion

thermoﬁsher.com

29 October 2019

Thermo Fisher Scientific VetMAX AIV Reagents Mode d'emploi

Marque: Thermo Fisher Scientific

Modèle: VetMAX AIV Reagents

Catégorie: Kits de voiture

Taper: Mode d'emploi

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