Saliva DNA Isolation Kit: High-Purity Non-Invasive Extraction
The Saliva DNA Isolation Kit (RU45400) provides a rapid and reliable method for purifying high-quality genomic DNA from both fresh and preserved saliva samples. As saliva becomes an increasingly preferred alternative to blood for non-invasive sample collection, researchers require a robust extraction method that effectively removes PCR inhibitors and stabilisers found in preservation buffers. This kit addresses these challenges, delivering DNA of the highest integrity suitable for sensitive downstream applications in diagnostics, epidemiology, and population studies.
Utilising advanced spin-column chromatography, the kit processes samples efficiently without the use of hazardous phenol or chloroform. The protocol involves a simple lysis step with Proteinase K, followed by binding of the DNA to the spin column matrix. Impurities, salts, and proteins are washed away, and the purified genomic DNA is eluted in a flexible volume. The system is fully compatible with samples collected using Geneflow’s Saliva DNA Collection and Preservation Devices, ensuring a seamless workflow from collection to analysis.
Key Benefits:
- Rapid Protocol: Streamlined spin-column format. Isolate genomic DNA from up to 10 samples in under 30 minutes, maximising laboratory throughput.
- Versatile Compatibility: Process fresh or preserved samples. Optimised for use with both fresh saliva and samples stored in preservation buffers (e.g., Norgen Saliva DNA Preservative).
- High Purity & Yield: NGS-ready quality. Consistently yields DNA with an OD260/280 ratio of 1.7–2.1, ensuring success in sensitive applications like Next-Generation Sequencing (NGS) and qPCR.
- Non-Invasive Solution: Alternative to blood collection. Ideal for recruiting donors for large-scale studies where blood draw is impractical, offering painless collection without sacrificing DNA quality.
Technical Insights
Figure 1. High Quality and Yield of DNA from Saliva Samples.
Total DNA was isolated from 250 µL of eight different fresh saliva samples using Norgen’s Saliva DNA Isolation Kit (Lanes 1-8). For evaluation, 10 µL of each 100 µL DNA elution was run on a 1.2 % agarose gel. Note the high yield and quality of the DNA in all lanes. Lane M: Norgen’s Fast runner 1kb DNA Ladder.
Figure 2. Real-time PCR Consistency from Saliva Samples.
Norgen’s Saliva DNA Isolation Kit was used to isolate DNA from 250 µL samples of fresh saliva. Ten samples were used from same donor. From the 100 µL elution, 5 µL of saliva DNA was directly mixed to real-time PCR master mix (total reaction volume 20 µL), and the real-time PCR reaction was performed. The GAPDH gene was successfully amplified from the different samples without any inhibition and at the same Ct, indicating the isolation consistency and the excellent quality of saliva DNA for downstream applications.
Figure 3. Illumina MiSeq 16s rRNA metagenomics data from saliva samples preserved for over 6 years using Norgen’s Saliva DNA Collection and Preservation Devices.
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 6 years at room temperature. A) Principal Coordinate Analysis of 12 saliva microbiomes showing differences in the distribution of taxonomic classifications between samples up to kingdom level. B) Hierarchical clustering of 12 saliva microbiomes based on genus-level classifications, including a bar chart showing the relative abundance of genus-level classifications for each sample in the dendrogram.
Specifications
- Input Volume: Up to 0.5 mL preserved saliva or 0.25 mL fresh saliva
- Average Yield: 3–7 µg (from 0.25 mL fresh saliva, donor dependent)
- Purity: OD260/280: 1.7 – 2.1
- Elution Volume: 50 µL – 100 µL
- Time to Complete: 30 minutes (per 10 purifications)
- Format: Spin Column (Microcentrifuge)
- Shelf Life: 1 year (Proteinase K stable at room temperature or 4°C)