In this tutorial, you will learn how to use Norgen Biotek’s Saliva/Swab RNA Purification Kit (Cat. 69100), which provides a rapid method for the purification of total RNA from non-preserved saliva and nasal/throat swabs, and from preserved saliva collected on Norgen’s Saliva RNA Collection and Preservation Devices (Cat. RU53800) or preserved swabs collected in Norgen’s Total Nucleic Acid Preservative Tubes (Cat. 69200).

To learn more about this kit, please visit our website [ Ссылка ]

In this kit, you will find:
A detailed product insert, Mini Spin Columns, Collection Tubes, Elution Tubes, Wash Solution A, Solution WN, Lysis Buffer A, and Elution Solution A.

Lysate Preparation from Preserved Saliva Sample
- Transfer 250 µL of preserved saliva sample into a 2 mL tube. Add 1x PBS pH 7.4 to make up the volume to 400 µL.
- Add 400 µL of Lysis Buffer A directly to the previous mix and mix by vortexing for 10 seconds.
- Add 400 µL of 96-100% Ethanol (provided by the user) to the lysate and mix by vortexing for 10 seconds.

Total RNA Purification
- Assemble a spin column with one of the provided collection tubes.
- Apply up to 600 µL of the lysate with the ethanol onto the column and centrifuge for 1 minute at 20,800 × g (~14,000 RPM).
- Discard the flow-through and reassemble the spin column with its collection tube.
- Depending on your lysate volume, repeat these steps as necessary to bind the remaining lysate volume.

- Apply 400 µL of Solution WN to the column and centrifuge for 1 minute at 20,800 × g (~14,000 RPM).
- Discard the flow-through and reassemble the spin column with its collection tube.
- Apply 400 µL of Wash Solution A to the column and centrifuge for 1 minute at 20,800 × g (~14,000 RPM).
- Discard the flow-through and reassemble the spin column with its collection tube.
- Wash the column a second time with Wash Solution A.
- Spin the column for 2 minutes to thoroughly dry the resin and discard the collection tube.

- Place the column into a fresh 1.7 mL Elution Tube provided with the kit.
- Add 50 µL of Elution Solution A to the column and centrifuge for 2 minutes at 420 × g (~2,000 RPM), followed by 1 minute at 20,800 × g (~14,000 RPM).

The purified RNA sample may be stored at -20°C for a few days. It is recommended that samples be placed at -70°C for long-term storage.

The purified RNA can be used for various downstream applications, such as RT-qPCR, NGS, Viral Detection and more!

To learn more about this kit, please visit our website [ Ссылка ]

Or, give us a call at 1-866-NORGENB (Toll-free) or email our product specialists at info@norgenbiotek.com.

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