Thermo Fisher Scientific SalivaReady Quick Start Guide

SalivaReady™ Solution

C

atalog Number A50608

Pub. No. MAN0019847 Rev. A.0

QUICK REFERENCE

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

clothing, and gloves. Safety Data Sheets (SDSs) are available from thermofisher.com/support.

ear appropriate protective eyewear,

Product description

SalivaReady™ Solution pr

epares saliva samples for real-time RT-PCR without the need for nucleic acid extraction or purification.

Contents and storage

Table 1 SalivaReady™ Solution

Cat. No.

[1]

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

C

[1]

A50608 2,000 40 mL –30°C to –10°C

Number of reactions Amount Storage

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

Equipment

Laboratory freezer

–30°C to –10°C

•

BSL-2 biological safety cabinet MLS

Centrifuge, with a rotor that accommodates 96-well plates MLS

Laboratory mixer, vortex or equivalent MLS

Single or multichannel pipettor (capable of pipetting 20 µL) MLS

ell Thermal Cycler, 0.2-mL block A24811, or equivalent

96‑W

Reagents and consumables

70% Isopropanol spray or wipes MLS

Reservoir for multichannel pipette, if needed MLS

MicroAmp™ Optical 96‑Well Reaction Plate with Barcode, 0.2 mL 4306737, or equivalent

MicroAmp™ Clear Adhesive Film 4306311

MicroAmp™ Adhesive Film Applicat

Sterile aerosol barrier (filtered) pipette tips thermofisher.com/pipettetips

or 4333183

MLS

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

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.

• Limit freeze-thaw cycles of SalivaReady™ Solution and saliva samples.

Prepare saliva samples

WARNING! Saliva samples have the pot

personal protective equipment (PPE) and handling samples in a BSL‑2 biological safety cabinet.

ential to transmit infectious diseases. Use safe laboratory procedures, including wearing

Before you begin

•

If the raw saliva samples are frozen, thaw completely at room temperature before processing.

• Thaw the SalivaReady™ Solution at room temperature, then mix by inversion at least 10 times.

(Optional) Heat inactivation of saliva samples

Heat inactivate the raw saliva sample before opening the sample container, if needed.

1. Heat the saliva samples at 95°C for 5 minutes.

2. Allow the samples to equilibrate to room temperature.

Prepare the samples

1. For the required number of samples, add 20 µL of SalivaReady™ Solution to each well of a new 96-well plate at room temperature.

2. Vortex each saliva sample at maximum speed for a minimum of 10 seconds, or until the sample appears homogenous.

Note: Samples that are particularly viscous or contain high amounts of particulate may require longer vortex times. Some samples

may contain particulate that does not fully homogenize.

3. Add 20 µL of saliva sample to each well containing the SalivaReady™ Solution, then pipet up and down to mix.

4. Seal the plate with a clear adhesive film, then vortex at maximum speed on all four sides along the skirt of the plate for 5 seconds

per side.

5. Centrifuge the plate for 30 seconds at 200 x g to collect the samples at the bottom of the wells.

6. Heat the plate in a thermal cycler using the following thermal conditions.

Step Temperature Time

1 62°C 5 minutes

2 92°C 5 minutes

3 4°C Hold

If necessary, the sealed plate may be stored at 4°C until used for real-time RT-PCR.

Not

e: We recommend using 7 µL of prepared sample in a 10 µL real-time RT-PCR reaction.

2 SalivaReady

™

Solution Quick Refer

ence