Thermo Fisher Scientific TaqCheck Quick Reference

QUICK REFERENCE

TaqCheck™ SARS-CoV

-2 Fast PCR Assay

Catalog Numbers A47693

Pub. No. MAN0019744 Rev. B.0

Note: For safety and biohazard guidelines, see the “Safety” appendix in the TaqCheck™ SARS-CoV-2 Fast PCR Assay User Guide (Pub. No. MAN0019745). Read the Safety Data Sheets (SDSs) and follow the handling instructions. Wear appropriate protective eyewear, clothing, and gloves.

Product description

The TaqCheck™ SARS-CoV-2 Fast PCR Assay is a multiplex real‑time RT‑PCR assay for the detection of SARS-CoV-2 viral RNA in human raw saliva samples. The assay has a multi-target design that compensates for emerging SARS-CoV-2 variants and mutations to provide conﬁdence in results. The assay contains primer and probe sets speciﬁc to the following targets:

Table 1 Assay targets, dyes, and quenchers

Target Dye Quencher

SARS-CoV-2 N gene

SARS-CoV-2 S gene

Human RNase P RPP30 gene

[1]

erves as an internal positive control to monitor sample quality.

The assay requires the following components:

• TaqCheck™ SARS‑CoV‑2 Control—RNA control that contains SARS-CoV-2 N protein and S protein target regions

• TaqCheck™ SARS‑CoV‑2 Control Dilution Buer—Dilution buer for the control

• TaqPath™ 1-Step RT-qPCR Master Mix, CG

For catalog numbers and storage conditions, see “Contents and storage”.

[1]

VIC™ dye QSY™ quencher

FAM™ dye QSY™ quencher

IMPORTANT! It is the r

own experimental design and analysis parameters.

esponsibility of the laboratories using the TaqCheck™ SARS-CoV-2 Fast PCR Assay to design and validate their

Contents and storage

The it

ems listed in the following table are required for the TaqCheck™ SARS-CoV-2 Fast PCR Assay. The items listed are sucient for

1,200 reactions.

Kit or product Cat. No. Amount Storage

TaqCheck™ SARS-CoV

TaqCheck™ SARS‑CoV‑2 Control 956127 3 × 10 µL ≤ –70°C

TaqCheck™ SARS‑CoV‑2 Control Dilution Buer A50486 3 × 250 µL –30°C to –10°C

TaqPath™ 1-S

tep RT-qPCR Master Mix, CG

-2 Fast PCR Assay A47693 690 µL –30°C to –10°C

• A15299

• A15300

• 5 × 1 mL

1 × 10 mL

•

–30°C to –10°C

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

Required materials not supplied

Unless otherwise indicated, all materials are available through thermoﬁsher.com. "MLS" indicates that the material is available from

ﬁsherscientiﬁc.com or another major laboratory supplier.

Catalog numbers that appear as links open the web pages for those products.

Item Source

Real-time PCR instrument and software

An Applied Biosystems™ real-time PCR instrument compatible with the dyes listed in Table 1 on page 1.

The assay was tested with the following instruments:

• Applied Biosystems™ 7500 Fast Real‑Time PCR Instrument

• Applied Biosystems™ QuantStudio™ 5 Real‑Time PCR Instrument, 96‑well, 0.2-mL block

• Applied Biosystems™ QuantStudio™ 5 Real‑Time PCR Instrument, 384‑well block

• Applied Biosystems™ QuantStudio™ 7 Flex Real-Time PCR Instrument, 384‑well block

(Recommended) QuantS

tudio™ Design and Analysis Software v2.5 or later

[1]

Equipment

Contact your local sales oce

thermoﬁsher.com/qpcrsoftware

Laboratory freezers

• –30°C to –10°C

• ≤ –70°C

BSL-2 biological safety cabinet, such as Herasafe™ 2030i Class 2 A2 Biological Safety C

abinets

Centrifuge (capable of achieving 1,400 × g), such as Megafuge™ 8 Small Bencht

Centrifuge Series or Multifuge X4 Pro Centrifuge Series

• MLS

• thermoﬁsher.com/tsx

• MLS

• thermoﬁsher

.com

• MLS

• thermoﬁsher

.com

• MLS

Microcentrifuge, such as Pico™ 17 Micr

ocentrifuge

• thermoﬁsher

.com

• MLS

Laboratory mixer, vortex or equivalent, such as Digital Vortex Mixers

Single and multichannel adjustable pipettors (2.00 µL to 1,000.0 µL)

• thermoﬁsher

ww.thermoﬁsher.com/cliptip

• w

• thermoﬁsher.com/ﬁnnpipette

.com

Cold block (96‑well or 384‑well) or ice MLS

Heat block or water bath (capable of reaching 95°C), such as Touch Screen Dry Bath/Block Heat

Liquid handler (if needed for automation)

er or Precision™ General Purpose Baths

[2]

• MLS

• thermoﬁsher

.com

MLS

Kits and reagents

TBE Buer (T

ris-borate-EDTA) (10X) B52, or equivalent

Tween®-20 Surfact-Amps™ Detergent Solution 28320

• AM9938 (1 x 100 mL)

Nuclease-free Water (not DEPC-Treated)

• AM9932 (1 x 1,000 mL)

70% Isopropanol spray or wipes MLS

2 TaqCheck

™

SARS-CoV-2 Fast PCR Assay Quick Reference

Tubes, plates, and other consumables

Item Source

(Recommended) S

terile tube with leak‑proof, screw‑top lid for sample collection

IMPORTANT! Do not use tubes that contain preservative.

[3]

One of the following, or equivalent:

• AM12500

• 339650

• 14-959-49B (ﬁsherscientiﬁc.com)

Reservoir for multichannel pipettes MLS

Sterile aerosol barrier (ﬁlt

ered) pipette tips thermoﬁsher.com/pipettetips

96‑well plate (for preparing saliva samples, not for RT-PCR) AB0796, or equivalent

code)

MicroAmp™ F

ast Optical 96-Well Reaction Plate, 0.1 mL

• 4346906 (with bar

• 4366932 (with barcode)

• 4346907 (without barcode)

• 4306737 (with bar

code)

• 4326659 (with barcode)

MicroAmp™ Optical 96-W

ell Reaction Plate, 0.2 mL

• N8010560 (without barcode)

• 4316813 (without barcode)

• 4309849 (with bar

code)

• 4326270 (with barcode)

MicroAmp™ Optical 384-W

ell Reaction Plate

• 4343814 (with barcode)

• 4343370 (without barcode)

MicroAmp™ Clear Adhesive Film 4306311

MicroAmp™ Optical Adhesive Film 4311971, 4360954

MicroAmp™ Adhesive Film Applicat

or 4333183

Nonstick, RNase-free microcentrifuge tubes (1.5 mL and 2.0 mL) thermoﬁsher.com/microtubes

DNase and RNase‑free tubes for mixing reagents (capable of mixing 5 mL and 50 mL) thermoﬁsher.com

Nunc™ 1.8‑mL Externally-Threaded Universal Tubes 374502

Nalgene™ General Long-Term Storage Cryogenic Tubes, 0.2 mL 5000-0020

Nunc™ Biobanking and Cell Culture Cryogenic Tubes, 4.5 mL 337516

Sterilin™ Certiﬁed Universal Containers – RNase, DNase, human DNA and Pyrogen Free, 30 mL

[1]

Use of QuantStudio™ Design and Analysis Software v2.5 is recommended, but not required. It is the responsibility of the laboratories using the assay to design and validate their own experimental design and analysis parameters.

[2]

Follow the guidelines provided by the manufacturer.

[3]

The use of the tubes listed in the table is recommended, but not required. Laboratories are responsible for validating their sample collection and preparation procedures for use with the assay.

• 30APPRN (Unlabelled)

• 30BPPRN (Graduated label)

General laboratory recommendations

• Implement standard operating procedures in your laboratory to prevent contamination, such as the following: – Frequent glove changes

– Frequent decontamination of surfaces, equipment, and pipettes with 10% bleach or decontamination solution, followed by 70%

ethanol

– Use of ultraviolet light during biosafety cabinet decontamination (when available)

• Saliva samples should always be treated as if infectious and/or biohazardous in accordance with safe laboratory procedures.

• To prevent degradation, keep master mixes, assays, and controls on ice or in cold blocks while in use. Limit freeze-thaw cycles.

• Aliquot reagents to prevent stock contamination and reduce the number of freeze-thaw cycles.

• To ensure reliable performance of the real‑time PCR instrument, perform preventive maintenance according to the instructions

provided by the manufacturer in the instrument documentation (see “Related documentation” on page 9).

TaqCheck™ SARS-CoV

-2 Fast PCR Assay Quick Reference 3

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