Overview

Norgen's Saliva DNA Collection and Preservation Devices are designed for 1) simple and non-invasive saliva collection and 2) preservation of DNA in saliva samples at ambient temperature. Each of the 50 Saliva DNA Collection and Preservation Devices consists of 3 components:

• Saliva Collection Funnel and Collection Tube
• Collection Tube Cap
• Norgen's Saliva DNA Preservative contained within a sealed squeezable ampoule

Saliva samples are collected by spitting inside the Collection Funnel which has been assembled with the Collection Tube. After collecting the required volume of saliva the Collection Funnel is removed and the contents of the Preservative Ampoule are then added and mixed with the collected saliva. The Saliva Collection Tube is subsequently sent to the laboratory for DNA isolation and analysis. DNA can be isolated from the preserved saliva samples using Norgen's Saliva DNA Isolation Kit (Cat. RU45400). Each of Norgen's Collection Tubes is labeled with a unique serial number that can be used for secure and anonymous tracking of the sample. The saliva DNA in preserved samples is stable for more than 2 years at room temperature. This kit is ideal for collecting and preserving DNA samples for epidemiological and population studies.

Saliva DNA Preservative

Norgen's Saliva DNA Preservative is an aqueous storage buffer designed for rapid cellular lysis and subsequent preservation of DNA from fresh specimens. The buffer prevents the growth of Gram-negative and Gram-positive bacteria and fungi, and also inactivates viruses allowing the resulting non-infectious samples to be handled and shipped safely. In addition, the buffer eliminates the need to immediately process or freeze samples and allows the samples to be shipped to centralized testing facilities at ambient temperature. The components of the buffer allow samples to be stored for more than 2 years without any detectable DNA degradation.

DNA Isolation from Preservative

Prior to saliva DNA isolation, vortex the Collection Tube containing preserved saliva for 10 seconds and incubate at 55°C for one hour. Saliva DNA can now be isolated from the preserved saliva samples using any commercially available method, including Norgen’s Saliva DNA Isolation Kit (Cat. RU45400), Norgen’s Saliva DNA Isolation 96-Well Kit (Cat. RU35200) and Norgen’s Saliva DNA Isolation Kit (Magnetic Bead System) (Cat. RU55400, RU62900). Also, Norgen’s Saliva DNA Isolation Reagent Kit (up to 4 mL) (Cat. RU35720) allows for DNA isolation from the preserved saliva samples using an alcohol precipitation method. The purified DNA is of the highest quality and can be used in a number of downstream applications including PCR, NGS and microarray analysis.

Details

Supporting Data

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Figure 1. Stability of DNA Preserved in Norgen's Saliva DNA Preservative at Room Temperature for over 2 Years. Saliva samples were collected from numerous donors and mixed, and then an equal volume of Norgen's Saliva DNA Preservative was added to the saliva. The preserved saliva was then stored at room temperature for up to 24 months. Saliva DNA was subsequently isolated at 4 months, 8 months, 16 months and 24 months from 0.5 mL of the saliva/preservative sample using Norgen's Saliva DNA Isolation Kit (Cat. RU45400). For visual analysis, 100 µL of the purified DNA was run on an agarose TAE gel. As it can be seen, there is no evidence of DNA degradation after the saliva samples are stored for 24 months at ambient temperatures in Norgen's Saliva DNA Preservative. Furthermore, the size of the DNA was maintained over 24 kb for the entire 24 month period. M: Norgen's UltraRanger 1 Kb DNA Ladder (Cat. 12100).

Figure 2. GAPDH Real Time PCR Amplification of Saliva DNA from Saliva Samples Stored at Room Temperature from Day 0 up to 24 Months. All isolated saliva DNA samples were successfully amplified without any sign of PCR inhibition and difference in Ct. Red: Day 0, Blue: 4 months, Purple: 8 months, Green: 16 months and Sky blue: 24 months. Black: No template control.

Figure 3. V3-V4 16s rRNA PCR amplification for Illumina MiSeq 16s rRNA library preparation. The saliva DNA was isolated using Norgen's Saliva DNA Isolation Kit (Cat. RU45400) from saliva that had been preserved for various periods of time up to 2 years at room temperature in Norgen’s Saliva DNA Preservative. The PCR result shows the clean PCR amplification at ~550 bp indicating the right target of V3-V4 16s rRNA , with no difference between day 0 and 24 months. M: FastRunner DNA Ladder (Cat. 12800).

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