Saliva DNA Isolation Kit

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.

Figure 4. Consistent DNA Isolation. Saliva samples were collected from 10 donors using Norgen's Saliva DNA Collection and Preservation Devices (Cat# RU49000). Norgen's Saliva Preservative was added to the saliva samples, and the 10 samples were then pooled together. Total DNA was then isolated from aliquots of the pooled and preserved saliva using columns from Norgen's Saliva DNA Isolation Kit (Cat# RU45400) or Norgen's Saliva DNA Isolation 96-Well Kit. Ten µL of each 100 µL elution from the 96-well plates were loaded on 1.2 % TAE agarose gel (Lanes 1 to 20), with the results indicating that the DNA isolation was consistent between the samples. S: DNA isolated using a single column (eluted in 200 µL). Lane M: Norgen's HighRanger 1kb DNA Ladder.

Figure 5. Real-time PCR Consistency from Saliva Samples. Saliva DNA was isolated from pooled saliva samples using Norgen's Saliva DNA Isolation 96-Well Kit and Norgen's Saliva DNA Isolation Kit (Cat# RU45400). From the 100 µL elution, 2 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, indicating the isolation consistency and the excellent quality of saliva DNA for downstream applications. Red: DNA samples isolated from the 96 well plate. Blue: DNA samples isolated using the single column. Black: Negative control.

Figure 6. Resolution of DNA Isolated from Preserved Saliva Samples. DNA was isolated from 0.5 mL of preserved saliva samples collected from 6 different donors (A to F) using Norgen's Saliva DNA Isolation Kit (Magnetic Bead System). As a control, DNA was isolated from one preserved saliva sample using Norgen's Saliva DNA Isolation Kit (column format, Cat. RU45400). For evaluation, 10 µL of each 75 µL elution was run on a 1X TAE 1.2% agarose gel. Similar DNA integrity and yield were observed between the two Norgen methods, indicating the robust performance of the Saliva DNA Isolation Kit (Magnetic Bead System). Marker = Norgen's HighRanger DNA Ladder.

Figure 7. Comparison of Saliva DNA Isolation Methods (Column vs. Magnetic Bead System). DNA was isolated from 0.5 mL of preserved saliva samples collected from 6 different donors (A to F) using Norgen's Saliva DNA Isolation Kit (Magnetic Bead System). As a control, DNA was isolated from one preserved saliva sample using Norgen's Saliva DNA Isolation Kit (column format, Cat. RU45400) . All DNA purified using Norgen’s Saliva DNA Isolation Kit (Magnetic Bead System) [A to F] showed a comparable concentration to DNA isolated using Norgen’s Saliva DNA Isolation Kit (column format, Cat. RU45400), indicating the consistent and robust performance of the Saliva DNA Isolation Kit (Magnetic Bead System).

Figure 8. High Quality DNA Confirmed by Real-time PCR. DNA was isolated from 0.5 mL of preserved saliva samples collected from 6 different donors (A to F) using Norgen's Saliva DNA Isolation Kit (Magnetic Bead System). As a control, DNA was isolated from one preserved saliva sample using Norgen's Saliva DNA Isolation Kit (column format, Cat. RU45400). Saliva DNA quality was confirmed by Real-time PCR using 8 µL of saliva DNA (total PCR reaction volume was 20 µL) to detect GAPDH. GAPDH was detected from all the saliva DNA isolated using the Magnetic Bead System (blue lines), similar to Norgen’s column based method (Saliva DNA Isolation Kit; Cat. RU45400) (red line). The successful GAPDH detection without PCR inhibition indicates the excellent saliva DNA quality from Norgen’s Saliva DNA Isolation Kit (Magnetic Bead System).

Figure 9. Compatibility Test with a Competitor’s Saliva Collection Device. Identical saliva samples were equally divided into Norgen’s Saliva DNA Collection and Preservation Devices (Cat. RU49000) and Competitor D’s saliva collection device. DNA was then isolated from 0.5 mL of preserved saliva using Norgen’s Saliva DNA Isolation Kit (Magnetic Bead System). For evaluation, 10 µL from each 75 µL of elution were run on 1X TAE 1.2% agarose gel. For the real-time PCR, 8 µL of DNA elution was add to total 20 µL of PCR reaction to detect GAPDH (Blue: Norgen, Red: Competitor D). Marker = Norgen’s HighRanger DNA Ladder. The results show that Norgen's kit was able to isolate high yields of pure DNA from both collection devices, indicating the robust performance of the kit.

Figure 10. Resolution of DNA Isolated from Preserved Saliva Samples from Different Donors. DNA was isolated from 0.5 mL of preserved saliva samples collected from 8 different healthy donors using Norgen's Saliva DNA Isolation 96-Well Kit (Magnetic Bead System). For evaluation, 10 µL of each 50 µL elution was run on a 1X TAE 1.2% agarose gel. Marker = Norgen's HighRanger DNA Ladder (Cat. 11900).

Figure 11. High Quality DNA Confirmed by Real-time PCR. DNA was isolated from 0.5 mL of preserved saliva samples collected using Norgen's Saliva DNA Isolation 96-Well Kit (Magnetic Bead System). Saliva DNA quality was confirmed by Real-time PCR using 2 µL of saliva DNA (total PCR reaction volume was 20 µL) to detect GAPDH. GAPDH was detected from all the saliva DNA isolated using the Saliva DNA Isolation 96-Well Kit (Magnetic Bead System). The successful GAPDH detection without PCR inhibition indicates the excellent saliva DNA quality from Norgen’s Saliva DNA Isolation 96-Well Kit (Magnetic Bead System).